Compositions and methods for the treatment of presbyopia

ABSTRACT

The invention provides compositions and methods for the treatment of presbyopia. The compositions comprise aceclidine hydrochloride, polysorbate 80, mannitol, anhydrous hypromellose, disodium edetate dihydrate, benzalkonium chloride, sorbic acid and anhydrous sodium citrate and optionally tropicamide.

CROSS-REFERENCE

This application is a continuation of U.S. application Ser. No.17/502,066, Oct. 15, 2021, which is a continuation of U.S. applicationSer. No. 16/747,070, filed Jan. 20, 2020, now Issued U.S. Pat. No.11,179,328, which is a division of U.S. application Ser. No. 16/106,730,filed Aug. 21, 2018, now Issued U.S. Pat. No. 11,179,327, which is acontinuation in part of U.S. application Ser. No. 15/956,936, filed Apr.19, 2018, which is a continuation in part of U.S. application Ser. No.15/864,703, filed Jan. 8, 2018, now Issued U.S. Pat. No. 10,617,763,which is a continuation in part of U.S. application Ser. No. 15/235,431,filed Aug. 12, 2016, now Issued U.S. Pat. No. 10,052,313, which is acontinuation in part of U.S. application Ser. No. 15/073,139, filed Mar.17, 2016, now Issued U.S. Pat. No. 9,833,441, which is a continuation inpart of U.S. application Ser. No. 15/073,089, filed Mar. 17, 2016, nowIssued U.S. Pat. No. 9,844,537, which is a continuation in part of U.S.application Ser. No. 14/742,903, filed Jun. 18, 2015, now issued as U.S.Pat. No. 9,320,709, each of which are incorporated by reference hereinin their entireties.

BACKGROUND OF THE INVENTION

As a person ages the minimum distance from the eye at which an objectwill come into focus, provided distance vision is corrected or isexcellent unaided, increases. For example, a 10-year-old can focus on anobject or a “focal point” only three inches (0.072 meters) from theireye while still retaining excellent distance vision; a 40-year-old atsix inches (0.15 meters); and a 60-year-old at an inconvenient 39 inches(1.0 meter). This condition of increasing minimum focal length inindividuals with excellent unaided distance vision is called presbyopia,loosely translated as “old-man eye”.

Excellent unaided distance vision is also known as emmetropia. Theinability to focus on distant focal points is known as myopia and theinability to focus on near focal points is known as hyperopia.Specifically, “distance” vision is considered any focal point 1 meter ormore from the eye and near vision is any focal point less than 1 meterfrom the eye. The minimum focal length at which an object will come intofocus is known as the “near point”. The change in focus from distance tothe near point and any focal point in between is called accommodation.Accommodation is often measured in diopters. Diopters are calculated bytaking the reciprocal of the focal length (in meters). For example, thedecrease in accommodation from a 10-year-old eye to a 60-year-old eye isabout 13 diopters (1÷0.072 meters=13.89 diopters; 1÷1 meter=1 diopter).

The highest incidence of first complaint of presbyopia occurs in peopleages 42-44.

Presbyopia occurs because as a person ages the eye's accommodativeability which uses near reflex-pupil constriction, convergence of theeyes and particularly ciliary muscle contraction, decreases. Thisreduction in accommodation results in an inadequate change in the normalthickening and increased curvature of the anterior surface of the lensthat is necessary for the shift in focus from distant objects to nearobjects. Important near focus tasks affected by presbyopia includeviewing computer screens (21 inches) and reading print (16 inches).

Presbyopia is a normal and inevitable effect of ageing and is the firstunmistakable sign for many in their forties that they are getting older.One study found that more than 1 billion people worldwide werepresbyopic in 2005. This same study predicted that number to almostdouble by the year 2050. If everyone over the age of 45 is considered tobe presbyopic, then an estimated 122 million people in the United Statesalone had presbyopia in 2010. As baby boomers reach the critical age,this number is only going to increase.

Presbyopia carries with it a stigma resulting from the limitation inability to quickly function at many tasks requiring focusing at bothdistant and near points, which once occurred almost immediately. In thepresbyopic patient, these tasks can be performed only by the use ofeyeglasses, contact lenses or after undergoing invasive surgery. Onesuch optical modification, the monovision procedure, can be executedwith the use of glasses, contact lenses or even surgery. The monovisionprocedure corrects one eye for near focus and the other eye for distancefocus. However, monovision correction is normally accompanied by loss ofdepth perception and distance vision particularly in dim light (e.g.night). Other surgical procedures that have been developed to relievepresbyopia include: (1) the implantation of intraocular lenses(INTRACOR®; registered trademark of Technolas Perfect Vision GMBH); (2)reshaping of the cornea (PresbyLASIK and conductive keratoplasty); (3)scleral band expansion; and (4) implantation of corneal inlays (FlexivueMicrolens®; registered trademark of PresbiBio LLC, Kamra®; registeredtrademark of AcuFocus, Inc. and Vue+). Kamra® corneal inlaysmanufactured by AcuFocus work by inlaying a pinhole on the cornea toincrease the depth of focus.

A similar effect can be achieved with general miotic agents, such aspilocarpine (a non-selective muscarinic acetylcholine receptor agonist),carbachol (a non-selective muscarinic acetylcholine receptor agonist),and phospholine iodide (an acetylcholinesterase inhibitor). Thesegeneral miotics can induce a pinhole pupil at sufficient concentrationsto achieve pupils below 2.0 mm and potentially extend depth of focusmuch like an inlay, but at concentrations sufficient to cause pinholepupil diameters of 2.0 mm or less these agents trigger increased ciliarymuscle contraction and induce accommodation of any remaining reserves,improving near vision at the expense of distance vision in individualswho still retain some accommodative function. The side effects ofciliary spasm induced migraine like brow pain and blurred distancevision from induced myopia beyond the ability of a pinhole pupil tocorrect then necessitate using weaker concentrations with much shorteracting and more marginal effect, such as found with pilocarpine. In suchcases even slight hyperopia helps offset the induced myopia while evenvery small increments of myopia, which is very common, exacerbate it. Inextreme cases, such ciliary muscle spasms may possibly be associatedwith anterior chamber shallowing and pull on the or a serrata of theretina, resulting in a retinal tear and or retinal detachment.

Miotic agents have been described in various patent and patentapplications for the treatment of presbyopia. U.S. Pat. Nos. 6,291,466and 6,410,544 describe the use of pilocarpine to regulate thecontraction of ciliary muscles to restore the eye to its resting stateand potentially restore its accommodative abilities.

U.S. Pat. No. 8,524,758 describes the use of pilocarpine with thenon-steroidal anti-inflammatory, diclofenac, to reduce brow ache fromciliary spasm and increase the time in which the ciliary musclecontraction is regulated. International PCT Application PublicationWO/2013/041967 describes the use of pilocarpine with oxymetazoline ormeloxicam to temporarily overcome ocular conditions such as presbyopia.

U.S. Pat. No. 8,299,079 (HEK Development LLC) describes the use ofdirect acting general miotic agents such as pilocarpine, carbachol andphospholine iodide with the alpha 2 selective vasoconstrictorbrimonidine at a concentration from 0.05% to 3.0% w/v. However, the useof brimonidine concentrations of about 0.20% (or any at or above 0.05%)w/v induces ciliary spasm with often migraine intensity brow and/orheadaches, and frequently results in increased rebound hyperemia. Forexample, rebound redness occurs in 25% of patients using brimonidine0.20% w/v (Alphagan®, registered trademark of Allergan, Inc.) twicedaily.

US Patent Application Publication No. 2014/0113946 describes the use ofpilocarpine with the alpha 1 and mild alpha 2 agonist vasoconstrictoroxymetazoline, demonstrating limitations in distance sharpness andduration, whereby a cohort largely restricted to mild hyperopes isrequired to neutralize the induced myopia (Table 5). Of the 16 eyestreated only three were −0.25 to −0.50 diopters, and eight were mildlyhyperopic. Of the −0.50 diopter eyes two were reduced to 20.40 distance.Further, duration was limited as full effect became diminished in aboutfour hours. Pupil size range was from 2.0 mm to 2.7 mm, where enhancednear effect and distance sharpness from depth of focus was minimal toabsent.

These attempts at miotic treatment for presbyopia all induce transientmyopia of several diopters reducing distance vision to about legalblindness or worse at the expense of improved near vision for the fullduration of their action, typically lasting several hours. This myopiceffect is amplified by the exponential drop off in distance acuity witheven small increments of nominal myopia in terms of unaided untreatedvision. For example, a person having mild myopia (e.g. sphericalequivalents of −0.25 D, −0.50 D) that is usually associated with glassesfree distance vision, typically will have several lines of distancevision loss after instillation of pilocarpine 1% (i.e. sphericalequivalent of −0.75 D.).

Miotics historically used to treat glaucoma, other than pilocarpine,particularly aceclidine, are also associated with ciliary spasm, browand/or headache, and myopic blur.

Further, aceclidine is unstable in solution. Normally, aceclidine isstored in a two-bottle system; one bottle containing the lyophilizedaceclidine and the second bottle containing the diluent necessary toreconstitute the lyophilized aceclidine before topical instillation.This two container system poses the risk of improper reconstitution andtherefor improper treatment of presbyopia.

However, the primary issue with its use as a presbyopic miotic is theattendant pain and in some cases distance blur that may be induced.

U.S. Pat. No. 9,089,562 describes a composition containing aceclidinecombined with a cycloplegic agent, such that in preferred embodimentsaceclidine 1.45% is combined with tropicamide 0.042%. The addition ofthe cycloplegic agent at extremely low concentrations (less than 0.10%)surprisingly still results in pupil miosis and allows for usefuldistance and improved near vision without ciliary spasm (often amigraine like brow ache that can be extremely painful and disabling),which is induced by the use of aceclidine alone. Further, aceclidine andthe cycloplegic agent require particular narrowly defined ratios andranges of concentrations relative to each other such that complicationsin the manufacturing and regulatory process, particularly the need forlyophilization of aceclidine to allow its stable storage, and attendanteffects of cryoprotectant/lyoprotectant (hereinafter referred to as“cryoprotectant”) required, where it is a discovery of the presentinvention the addition of a cryoprotectant such as a polyol, in apreferred embodiment mannitol, results in reduced efficacy of thedefined ranges and ratios of concentrations of U.S. Pat. No. 9,089,562.Due to these medical and practical inefficiencies, it is discovered anaceclidine composition requiring same or slightly higher concentrationsof aceclidine and much lower concentrations than U.S. Pat. No. 9,089,562or in some cases no cycloplegic agent, while allowing for formulationmodifications to lyophilize aceclidine would be preferred for thetreatment of presbyopia with necessary commercially stable formulations.However, to date, no aceclidine composition with amounts of cycloplegicagent lower than that claimed in U.S. Pat. No. 9,089,562 has beeneffective to treat presbyopia because, as mentioned above, aceclidinealone, particularly young and middle-aged presbyopes (ages 45 to 58),severe ciliary spasms and may cause accommodative induced distance blurin some subjects.

Thus, there is a need in the art for a treatment of presbyopia that isnon-invasive and convenient with minimal side effects. Specifically,there is a need for an ophthalmological composition that will allow aperson suffering from presbyopia to focus on near objects withoutsignificant side effects such as diminished distance vision, blurredvision, pain, redness, impaired night driving or incapacitating dimlight vision, induced nasal congestion, or risk of retinal detachment.Further, there is a need in the art for storage stable aceclidinecompositions.

Further, there is a need in the art for a reduction or elimination ofthe need for a cycloplegic agent to be used with aceclidine potentiallyenhancing duration and efficacy, as well as for means of storage ofstable aceclidine compositions, where such compositions preferablyenhance both distance and near depth of focus allowing pupil miosis to a1.50 to 2.0 mm range without clinically significant side effects.

SUMMARY OF THE INVENTION

In certain embodiments, the present invention is directed tocompositions and methods for the treatment of presbyopia.

In certain embodiments, the present invention is directed tocompositions and methods for the treatment of presbyopia comprising amuscarinic agonist, wherein the muscarinic agonist preferentiallyactivates M1 and M3 muscarinic acetylcholine receptors. In still morepreferred embodiments the muscarinic agonist is more highly selectivefor M1 than M3. In certain embodiments, the present invention isdirected to compositions and methods for the treatment of presbyopiacomprising a muscarinic agonist that preferentially activates M1 and M3muscarinic acetylcholine receptors.

In certain embodiments, the present invention is directed tocompositions and methods for the treatment of presbyopia comprising amuscarinic agonist selected from the group consisting of aceclidine,talsaclidine, vedaclidine, sabcomeline, cevimeline, WAY-132983, AFB267B(NGX267), AC-42, AC-260584, 77-LH-28-1, and LY593039 or anypharmaceutically acceptable salts, esters, analogues, prodrugs orderivatives thereof.

In certain embodiments, the present invention is directed tocompositions and methods for the treatment of presbyopia comprising amuscarinic agonist that activates only M1 muscarinic acetylcholinereceptors.

In certain other embodiments, the present invention is directed to anophthalmological composition for the treatment of presbyopia comprisingaceclidine, preferably at a concentration from about 0.25% to about 2.5%w/v, more preferably from about 0.75% to about 2.5% w/v.

In certain other embodiments, the present invention is directed to anophthalmological composition for the treatment of presbyopia comprisingaceclidine and a selective α-2 adrenergic receptor agonist.

In certain preferred embodiments, the ophthalmological compositions ofthe present invention further comprise sodium chloride.

In certain preferred embodiments, the present invention is directed toophthalmological compositions for the treatment of presbyopia comprisingaceclidine, one or more nonionic surfactants and a viscosity agent.

In certain preferred embodiments, the one or more nonionic surfactantsof the present invention are selected from the group consisting of apolysorbate, tyloxapol, a poloxamer, a cyclodextrin, vitamin E TPGS anda polyoxyl castor oil, a polyoxyl stearate, polyethylene glycol alkylether and2-[[10,13-dimethyl-17-(6-methylheptan-2-yl)-2,3,4,7,8,9,11,12,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-3-yl]oxy]ethanol,more preferably a polysorbate and/or a cyclodextrin and even morepreferably polysorbate 80, poloxamer 407 and/or poloxamer 188,preferably at a total concentration from about 2% to about 7% w/v.

In certain preferred embodiments, the viscosity agent provides aviscosity of the total composition of about 25 centipoise (“cps”) ormore at 25° C. and 0 shear, preferably from about 50 to about 10,000cps, more preferably from about 100 to about 5,000 cps and mostpreferably from about 150 to about 450 cps.

In certain preferred embodiments, the viscosity agent is selected fromthe group consisting of a cellulose derivative, hyaluronate, a carbomerand a gum, more preferably high molecular weight carboxymethylcellulose, carbomer 940 and hydroxypropylmethyl cellulose.

In certain preferred embodiments, the ophthalmological compositions ofthe present invention further comprise one or more antioxidants selectedfrom the group consisting of ethylenediaminetetraacetic acid (EDTA),ethylenediaminetetraacetic acid dihydrate, sodium citrate and citratebuffer, preferably selected from the group consisting ofethylenediaminetetraacetic acid dihydrate and sodium citrate or citratebuffer.

In certain preferred embodiment, ophthalmological compositions of thepresent invention have a pH of about 4.0 to about 8.0.

In certain other embodiments, the present invention is directed to anophthalmological composition of the present invention comprising:

-   -   a general miotic agent, a muscarinic agonist or aceclidine;    -   optionally a cycloplegic agent;    -   optionally a selective α-2 adrenergic receptor agonist;    -   a viscosity enhancer; and    -   a surfactant selected from the group consisting of an anionic        surfactant, a nonionic surfactant,    -   and a combination thereof.

In certain embodiments, the present invention is directed tocompositions for the treatment of presbyopia comprising about 1.75% w/vaceclidine, about 4.0% w/v polysorbate 80, about 2.5% w/v mannitol,about 1.2% w/v hydroxypropylmethyl cellulose, about 0.1% w/vethylenediaminetetraacetic acid, about 0.02% w/v benzalkonium chloride,about 0.12% w/v potassium sorbate and about 0.077% w/v citrate, whereinthe composition has a pH of about 5.0.

In certain other embodiments, the present invention is directed tocompositions for the treatment of presbyopia comprising about 1.75% w/vaceclidine, about 4.0% w/v polysorbate 80, about 2.5% w/v mannitol,about 1.2% w/v hydroxypropylmethyl cellulose, about 0.1% w/vethylenediaminetetraacetic acid, about 0.02% w/v benzalkonium chloride,about 0.12% w/v potassium sorbate and about 0.1% w/v citrate, whereinthe composition has a pH of about 5.0.

In certain other embodiments, the present invention is directed tocompositions for the treatment of presbyopia comprising about 1.75% w/waceclidine hydrochloride, about 4.0% w/w polysorbate 80, about 2.5% w/wmannitol, about 1.25% w/w anhydrous hypromellose, about 0.1% w/vdisodium edetate dihydrate, about 0.02% w/w benzalkonium chloride, about0.12% w/w sorbic acid and about 0.077% w/w anhydrous sodium citrate andoptionally, 0.01% w/w tropicamide, wherein the composition has a pH ofabout 5.0.

In certain other embodiments, the present invention is directed tocompositions for the treatment of presbyopia comprising about 1.75% w/waceclidine hydrochloride, about 4.0% w/w polysorbate 80, about 2.5% w/wmannitol, about 1.25% w/w anhydrous hypromellose, about 0.1% w/wdisodium edetate dihydrate, about 0.02% w/w benzalkonium chloride, about0.12% w/w sorbic acid and about 0.1% w/w anhydrous sodium citrate andoptionally, 0.01% w/w tropicamide, wherein the composition has a pH ofabout 5.0.

In certain other embodiments, the present invention is directed tocompositions for the treatment of presbyopia comprising about 1.40% w/vaceclidine, about 2.0% w/v polyoxyl stearate, about 2.5% w/v mannitol,about 0.1% w/v ethylenediaminetetraacetic acid, about 0.02% w/vbenzalkonium chloride, about 0.12% w/v potassium sorbate and about 0.1%w/v citrate, wherein the composition has a pH of about 5.0.

In certain other embodiments, the present invention is directed to amethod of treating presbyopia comprising administering to a subject inneed thereof a composition of the present invention.

In certain other embodiments, the present invention is directed to amethod of treating a refractive error of the eye in a subject in needthereof comprising administering to a subject in need thereof apharmaceutically acceptable amount of a composition of the presentinvention, wherein the size of the pupil is reduced to from about 1.5 toabout 2.5 millimeters, preferably from about 1.7 to about 2.0millimeters, and wherein the refractive error of the eye is selectedfrom presbyopia, myopia, hyperopia, astigmatism or a combinationthereof.

The present invention is further directed to a method of increasing thevisual depth of field (i.e. depth of focus) comprising administering toa subject in need thereof a pharmaceutically effective amount of anophthalmological composition of the present invention.

The present invention is further directed to a method of increasing thevisual depth perception upon improving near vision unaided comprisingadministering to a subject in need thereof a pharmaceutically effectiveamount of an ophthalmological composition of the present invention inboth eyes (binocular vision), wherein such binocularity further enhancesnear vision beyond that of either eye separately.

The present invention is further directed to a method of improvingvision in a subject with ammetropia (vision abnormality), comprisingadministering to a subject in need thereof a pharmaceutically effectiveamount of a composition of the present invention.

The present invention is directed to methods of treating low regularastigmatism, low or high irregular astigmatism, keratoconic ectasia, andlow myopia, or hyperopia, with or without astigmatism, comprisingadministering to a subject in need thereof an ophthalmologicalcomposition of the present invention.

The present invention is further directed to a method for stabilizingthe composition of claim 1 comprising maintaining the composition at atemperature from about 2 to about 8° C.

The present invention is further directed to a method for stabilizing anaqueous aceclidine composition comprising the steps of:

-   -   adding a surfactant selected from polyoxyl 40 stearate, a gamma        cyclodextrin, sulfobutylether β-cyclodextrin, 2-hydroxypropyl        cyclodextrin, sodium lauryl sulfate, sodium ester lauryl        sulfate, a poloxamer, a polysorbate, sorbitan monolaurate,        sorbitan monopalmitate, sorbitan monostearate, sorbitan        monooleate, a polyoxyl alkyl, a cyclodextrin and combinations        thereof to the composition, preferably polyoxyl 40 stearate;    -   adding a tonicity adjustor selected from mannitol, sodium        chloride, potassium chloride, glycerin and combinations thereof,        preferably mannitol;    -   optionally adding a viscosity enhancer selected from the group        consisting of guar gum, hydroxypropyl-guar, xanthan gum,        alginate, chitosan, gelrite, hyauluronic acid, dextran, a        carbomer and combinations thereof to the composition, preferably        carbomer 940, buffering the pH of the composition to from about        4.0 to about 6.0, preferably 4.75; and maintaining the        composition at a temperature from about 2 to about 8° C.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is a graphical representation of the effects of pilocarpine andaceclidine with or without tropicamide and with or without a carrier onnear and distance vision in a patient over the age of 45.

FIG. 2 is a graphical representation of the effects of addition ofnon-ionic surfactants and viscosity agents on near vision acuity andduration of effect. Line-Hours denotes lines improved times duration ofeffect.

FIG. 3 is a graphical representation of the Efficacy Index for formulas#L33-#L94. Box color denotes a comfort level of good for white, fair forcross-hatched and poor for black.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is directed to compositions and methods oftreating presbyopia, irregular astigmatism, and/or refractive error,comprising administering to a patient in need thereof a pharmaceuticalcomposition comprising a muscarinic agonist that preferentiallyactivates M1 and M3 muscarinic acetylcholine receptors, preferablyactivate M1 more than M3 and most preferably aceclidine or itsderivatives. Aceclidine has been surprisingly and unexpectedlydiscovered to provide enhanced presbyopic reversal with negligible sideeffects day or night (when viewing includes one or more direct orreflected light sources).

Aceclidine is traditionally used as a treatment for glaucoma. Whenaceclidine is used to treat glaucoma, it is normally stored in atwo-bottle system; one bottle containing the lyophilized aceclidine andthe second bottle containing the diluent necessary to reconstitute thelyophilized aceclidine before topical instillation. Romano J. H.,Double-blind cross-over comparison of aceclidine and pilocarpine inopen-angle glaucoma, Brit J Ophthal, August 1970, 54(8), 510-521. It isa further aspect of the present invention to provide an aqueousaceclidine composition that is stable in combination with cold chainstorage. It is yet a further aspect of the present invention to providea method of stabilizing aqueous aceclidine by combining effectiveexcipients, pH ranges and temperature ranges.

The compositions and methods of the present invention treat presbyopiaby improving depth of focus in patients with presbyopia by administeringan ophthalmological composition to the eye that reduces pupil dilationin the dark or in dim light, produces a particular degree and durationof miosis without accommodation, provides cosmetic whitening and/orinduce redness prophylaxis. The compositions and methods of the presentinvention also do not cause significant pupil rebound, tachyphylaxis,ciliary spasms, induction of myopia or reduction in distance vision.Additionally, the compositions and methods of the present inventionallow for the further improvement in visual acuity and depth perceptionof binocular (both eyes) treatment.

The ophthalmological composition of the present invention surprisinglycreates a pupil of from about 1.5 to about 2.4 mm at the anterior irisplane and about 2.0 mm at the corneal surface. Not wishing to be held toparticular theory the clinical effect appears to involve both withmodulated increase in accommodative tone and enhanced pinhole near depthof focus for improved near vision, estimated to be about −1.25 D orless, but restricted in power to remain within the range of pinholecorrection for distance, found to be about −1.00 D or less creating asum increase that may in some cases create a near vision add of +2.00 Dor more without distance blur; and with a reduction or ablation of theredness that is otherwise a hallmark of the use of miotic agents. Thepupil miosis of the present invention with such modulation andrestriction of peak accommodative tone is superior to the pinhole effectof the Kamra® and Flexivue Microlens® corneal inlays, allowing binoculartreatment without peak dimming. Pupil miosis of the present inventionwith modulated accommodation is also superior to inlays because theconstriction of the actual pupil does not result in the attendant severenight vision disturbance caused by the light scattering borders of thepre-corneal pinholes created by the inlays. Further pupil miosisprovides a greater field of vision and transmission of more focusedlight, and in a discovered optimal pupil range of about 1.5 mm to 2.1 mmusing formulation discoveries of the present invention does so withnegligible to mild and very tolerable dimming and enhanced contrast,distance vision, reduced glare at night, and improved near vision.

The use of aceclidine has a minimal effect on the longitudinal ciliarymuscle, thus reducing risk of retinal detachment when compared to theuse of general muscarinic agonists such as pilocarpine and carbachol.The further inclusion of a cycloplegic agent resulted in only 0.04 mm ofanterior chamber shallowing. Aceclidine, particularly as enhanced forthe present invention, also has greater magnitude, duration, and controlof minimum pupil diameter than conventional pilocarpine with or withoutalpha agonists, and less anterior chamber inflammation with chronic use.Compositions of the present invention achieve these advantages byallowing both pinhole near vision depth perception benefit and modestaccommodative increase below the threshold of induced myopic distanceblur through the miotic pupil, whereby, not wishing to be held toparticular theory, it is believed the rate of miosis and the rate ofaccommodative increase maintain a synchronous balance in preferredembodiments allowing pinhole correction of otherwise inducedaccommodative blur in prior art applications of miotics for presbyopiccorrection. This combination thus is found to avoid the distance blurtypically seen in patients as a response to pilocarpine and/or carbacholinduced miosis without the formulation discoveries of the presentinvention, as well as the excessive accommodative myopia and ciliaryspasm manifested as brow ache or generalized migraine-like headache.

Such conventional formulations of pilocarpine, in order to affect anyreasonable duration of effect, are still restricted to less than orequal to about 4 hours in most cases, as the high ratio of accommodationto pupillary miosis requires minimal concentrations of pilocarpine ofabout 1.0% to minimize but not eliminate distance induced myopic blurand ciliary spasm. Further pilocarpine must be instilled monocularly tominimize intolerable distance blur to a still bothersome 2-3 lines ofdistance blur. Even instilled monocularly, pilocarpine still may createbothersome attendant distance blur and must be restricted to about 1.0%.Upon instillation of 1.0% pilocarpine pupil size is about 2.3 mm orlarger in most subjects and thereby restricts any significant pinholedepth perception benefit as well as any pinhole filtering of inducedmyopic rays. The restriction to about 1.0% for these conventionalformulations of pilocarpine with the attendant short duration and stillbothersome but reduced distance blur in emmetropes or myopes (somewhatneutralized in low hyperopes) are attempts to prevent extremely strongaccommodation of 5D to 11 D well known to occur at higher concentrationsof pilocarpine.

Any effects on accommodation may be further reduced or totallyeliminated in preferred embodiments by combining a miotic with acycloplegic agent in a narrow and particular ratio of miotic tocycloplegic, where such ratios as discovered for U.S. Pat. No.9,089,562, such as about 35:1 for a preferred embodiment, become greatlyincreased for the present invention in the presence of cryoprotectant asto a factor of about 300%-700%. Aceclidine is capable of producing theincreased depth of focus by both pupil miosis below 2.3 mm and modestaccommodation described in the present invention. Particularly enhancedmiosis occurs with use of compositions of the present invention. Thisenhanced miosis makes it possible to use an α-2 agonist at very lowconcentrations if desired to reduce mild eye redness. Other combinationsof inactive ingredients reduce or effectively eliminate induced rednesswithout such agonists. Further, due to the apparent and surprisinglyselective nature of aceclidine, and the commercially stable aceclidineformulation discoveries of the present invention, administration to theeye of compositions of the present invention result in a net stronglyenhanced near vision acuity from both pupil miotic pinhole effect andmoderate modulated ciliary accommodation. These beneficial effects areaccompanied by a filtering pupil effect, which eliminates any distanceblur from the accommodation, correcting residual refractive error andoptical aberrations as may exist to in many cases improve distancevision as well. Thus, the administration of aceclidine results in pupilmiosis without excessive accommodation and attendant distance blur.

However, aceclidine alone may cause substantial redness and brow ache insome individuals. Without formulation enhancement of the presentinvention such as requiring cycloplegic agent, cryoprotectant or both,aceclidine may produce either less than optimal pupil miosis at lowconcentrations or at higher concentrations require more than desiredpeak miosis to attain satisfactory duration of greater than 3-4 hours.However the use of a cycloplegic agent has been found to be highlysensitive to other inactive ingredients in the formulation not usuallyassociated with effects on active agents, and particularly forcryoprotectants as found to be preferred commercially for aceclidinereduce or eliminate the need for this cycloplegic requirement toextremely low concentrations in a preferred embodiment, rendering 0.042%sufficiently high when a cryoprotectant is present (e.g. a polyol suchas mannitol) to cause substantial loss of efficacy. Further, aceclidinewithout formulation enhancements of the present invention causes dimmingof vision in dim or absent lighting as well as ciliary pain above areasonably tolerable threshold that may last for an hour or more and besimilar to a severe migraine headache.

Certain embodiments of the present invention enhance the discoveredpreferred degree of pupillary miosis by providing a consistent range ofeffect of about 1.50-2.20 mm for most patients using a preferredembodiment of a nonionic surfactant and viscosity agent. Similar benefitmay be achieved using other permeation enhancers, particularlyhydroxypropylmethyl cellulose, high viscosity carboxymethyl cellulose,Carbopol® (polyacrylic acid or carbomer), and various viscosityadditives that increase drug residence time, such as xanthan gums, guargum, alginate, and other in situ gels well known to experts in the art.It is well known to experts in the art that the exact concentration of aspecific viscosity agent will depend on both the molecular weight forthat agent selected and the concentration, such that for increasedmolecular weight a reduced concentration can have the same viscosity.The present invention further prevents nasal congestion otherwiseoccurring when substantial aceclidine levels reach the nasal mucosa, dueto the rheologic properties of the preferred embodiment.

The combination of aceclidine and a low concentration of a selective α-2adrenergic receptor agonist (α-2 agonist or α-2 adrenergic agonist),such as fadolmidine, brimonidine or guanfacine, allows for the desiredmiotic effect with diminished or no redness. The use of lowconcentrations of a selective α-2 agonist results in substantialreduction of hyperemia with greatly reduced risk of rebound hyperemiathat is found in concentrations of about 0.06% w/v or more. Furthermore,the use of low concentrations of selective α-2 agonist does notadversely modify the pupil constriction caused by aceclidine. Incontrast, the use of brimonidine 0.20% w/v, when topically applied forpupil modulation for night vision, result in tachyphylaxis of pupilmodulation due to α-2 receptor upregulation in almost 100% of treatedsubjects within four weeks of use.

Unexpectedly, the addition of a cycloplegic agent results in reductionof any brow ache or associated discomfort by further reducing the degreeof ciliary spasms on topical instillation without impairing the mioticresponse. More unexpectedly and surprisingly, the ratio of 1.40%aceclidine to about 0.040% tropicamide in a preferred embodiment of U.S.Pat. No. 9,089,562 (35:1) becomes about 1.75% aceclidine to about 0.004%to 0.010% tropicamide (350:1, 175:1 respectively) in the presence ofmannitol, where 2.5% provides better effect than 4.0%.

The lack of impairment of the miotic response is an unexpectedsurprising discovery, as particular cycloplegic agents, such astropicamide, have known pupil dilating effects at concentrations as lowas 0.01% w/v (Grünberger J. et al., The pupillary response test as amethod to differentiate various types of dementia, Neuropsychiatr, 2009,23(1), pg 57). More specifically cycloplegic agents cause pupilmydriasis (i.e. dilation of the radial muscle of the iris). Further, theaddition of a cycloplegic agent to the miotic agent unexpectedlyincreases the time at which the pupil maintains the desired size rangewithout becoming too restricted. Peak miotic effect at 30-60 minutes canbe titrated in inverse relation to the cycloplegic concentration. Theconcentrations of tropicamide discovered in the present inventionapparently cause more relaxation of the ciliary muscle than the irisradial musculature. In fact, iris mydriasis is discovered to besuppressed by the addition of tropicamide to compositions containingconcentrations of aceclidine used in the present invention, with insteada more consistent level of miosis for the duration of the miotic effect.Additionally, and quite surprisingly, unexpectedly, and beneficially theaddition of tropicamide can reduce the degree of peak pupil miosiswithout inducing mydriasis thereby creating a more constant and idealpupil size throughout the drug induced miosis. This more consistentpupil size allows for beneficial near and distance vision without theadverse dimming or loss of resolution due to diffraction limits at thevery reduced pupil sizes seen at peak pupil miosis (e.g. 1.25 mm).

Previously, in U.S. Pat. No. 9,089,562, it was surprisingly found thatthe addition of at least 0.04% w/v cycloplegic agent resulted in anabatement of ciliary side effects caused by the administration ofaceclidine (1.40%) to the eye, in a preferred embodiment, but suchformulations are not as constituted sufficiently stable for commercialuse, and typically have a duration of about five to six hours maximum.

Several additional discoveries of the present invention allow forcommercially stable aceclidine formulations with enhanced efficacy andduration:

-   -   Equally or more surprising than the synergistic effects of        cycloplegics of 0.040% added to aceclidine 1.40%, is the        discovery of the present invention that combination of        aceclidine 1.50%-2.0%, and preferably about 1.75% and a        cryoprotectant, preferably a polyol, in a preferred embodiment        mannitol, particularly at 0.5% to 4.0% and most preferably about        2.5%, can achieve a similar pupil range with reduced or absent        ciliary side effects. The cryoprotectant when combined with        aceclidine can then be combined to allow lyophilization without        degradation of aceclidine and simultaneously further reduce or        eliminate the need for a cycloplegic agent for the present        invention vs. the teachings of cycloplegic concentration ranges        required in U.S. Pat. No. 9,089,562. Optionally, the addition of        a cryoprotectant can therefore also be used to greatly reduce        (i.e. no more than 0.025% w/v cycloplegic agent, preferably        0.004% to 0.015% and most preferably 0.005% to 0.010%) the        concentration of cycloplegic required to further eliminate mild,        but potentially bothersome, ciliary side effects particularly in        younger presbyopes and further modulate pupil miosis over        aceclidine and a cryoprotectant combinations alone, reducing and        in most cases eliminating any bothersome peak concentration        dimming, as found in preferred embodiments of the present        invention. In preferred embodiments it is discovered that        aceclidine about 1.50%-2.0% and more preferably 1.75% and        mannitol about 0.5%-4.0% and more preferably 2.5% provide        optimal concentration combinations for the present invention,        that are necessary but not sufficient for about 3 lines of near        improvement and 5 or more hours duration desired for an        effective topical presbyopic composition, where additional        formulation discoveries can further enhance the desired clinical        near improvement magnitude and duration;    -   It is surprisingly discovered that adding a viscosity agent to        compositions described above only modestly improves magnitude        and duration, however when first adding a nonionic surfactant,        such as polyoxyl stearate or polysorbate 80, optimal        concentrations are discovered that provide greatly improved        magnitude and duration for the present invention, to which        viscosity may then provide added duration much more        substantially than when added alone. For polysorbate 80 or        polyoxyl 40 stearate concentrations of 1.0% to 10.0%, and more        preferably about 2.5% to 5.0% w/v have been found to be        beneficial.

When formulation improvements are combined, preferred embodiments suchas aceclidine 1.75%, mannitol 2.5%, and polysorbate 80 2.75% result.Viscosity agents such as high viscosity carboxymethyl cellulose (“CMC”)are surprisingly discovered to moderately enhance magnitude and greatlyenhance duration, unlike with formulations in a. above alone. Highmolecular weight CMC concentrations of 0.75% to 1.75%, and mostpreferably about 1.40%, or hydroxypropylmethyl cellulose (“HPMC”) atabout 0.25% to 2.0%, more preferably about 0.50% or 1.50%, and mostpreferably about 1.0% to 1.25%, when combined result now in about +3lines of near vision improvement or greater, at a duration of 5-10hours, at a mean of about 7 hours or greater vs. pilocarpine 1.0% ofabout less than 4 hours;

Not wishing to be held to particular theory citrate in combination withEDTA as a preferred embodiment buffer appears to 1) reduce redness; 2)enhance sorbate preservative shelf life, and in combination of the abovewith BAK 0.005% to 0.02% (0.02% preferred) further enhances near visionlines to about 4 lines and duration to about 8 to 12 hours.

In a preferred embodiment, compositions of the present invention furthercomprise sodium chloride, preferably at a concentration from about 0.5%to about 1.5% w/v, more preferably from about 0.65% to about 0.9% w/v.Optionally, sodium chloride may be substituted with boric acid,preferably at 0.35% or potassium borate, preferably at 0.47%;

Not wishing to be held to particular theory, it appears the addition ofnonionic surfactant at optimized concentration of about 2.0% to about7.0% enhances permeation of aceclidine into the eye, which may relate tooptimal micellar size particularly once of micromicellar or nanomicellarrange. This increased permeation coincides with the desirable increasein magnitude and duration and absent tropicamide but in the presence ofmannitol with slight increases in ciliary sensation and dimming.Therefore, in the presence of the combined formulation enhancementsabove, where a cycloplegic agent is no longer required, addition of anonionic surfactant at concentrations found to be preferred may befurther improved with much lower concentrations of a cycloplegic agentthan those found in U.S. Pat. No. 9,089,562, such as the use of about0.042% tropicamide with aceclidine 1.40%. For the present invention thenpreferred embodiments include aceclidine of about 1.75%, mannitol 2.5%,polysorbate 80 of about 2.5% to 5.0%, CMC of about 1.42%, or HPMC ofabout 1.8% and tropicamide of about 0.004%-0.010%, more preferably about0.005% to 0.007%, and most preferably about 0.005%-0.006%.

Micelle formation above the critical micellar concentration may allowfor micelles to spread across the tear film surface and spread at lowconcentrations to cover this surface, while at higher concentrationsthese micelles becoming increasingly contracted and “squeezed” along thesurface. Not wishing to be held to particular theory, it is believed atan optimal concentration a minimal micelle diameter is achieved beforesignificant multiple lamellae (layering) occurs. It is believed that atthe optimal concentration nanomicelles of about 100 to 250 nm along thesurface are achieved surrounding the highly charged and hydrophilicaceclidine, facilitating its penetration through the very lipophilicepithelium;

Not wishing to be held to particular theory the addition of BAK 0.02% tosorbate about 0.10%, EDTA about 0.10%, in a preferred composition ofaceclidine 1.75%, mannitol 2.5%, tropicamide 0.01%, and citrate buffer(1 to 100 mM 3-5 mM preferred) is above the BAK critical micellarconcentration. BAK, being a cationic surfactant, and BAK micelles,creating an ionic micellar gradient with +charge NH4+ quaternarynitrogen bring on the polar heads aggregating outside and lipophilicalkyl chain on the hydrophobic tails aggregating on the inside may causesignificant similar aceclidine alignment due to its dipole withquaternary NH3 nucleophilic or NH4 protonated nitrogens oriented alongthe outside polar heads and more hydrophobic carbonyls C═O alonghydrophobic BAK micellar tails these preventing, greatly reducing, ormoderately reducing collisions of any nonionic aceclidine molecules—thenucleophiles—which if oriented in solution such that randomly theycollide with another aceclidine carbonyl will result in chemicalconversion of that aceclidine via nucleophilic attack at its targetedcarbonyl, which can recur from such nucleophiles to other aceclidines sooriented repeatedly and cause loss of stability without such BAKorientation via 0.005% and preferably 0.01% to 0.02% most preferredmicelles. The concentration of such nonionic nucleophiles at a preferredpH in the preferred embodiment is relatively low, but the ability ofthese nonionic nucleophiles to destabilize adjacent aceclidinesrepeatedly without themselves degrading is otherwise high. The resultmay be improved potency for 1 month plus of a mixed solution once openedin a dual chamber bottle and mixing occurs of lyophilizedaceclidine/mannitol with the remainder of the formulation in the diluentand or improved stability sufficient for commercialization in solution,either at room temperature or via cold chain;

It is discovered that BAK alone does not provide sufficient bacterialand fungal preservative efficacy but that BAK and sorbate, or sorbatealone satisfactorily preserve diluent and or mixed solutions of theinvention;

Not to be wishing to be held to particular theory preferred embodimentsof the present invention such as containing 1.25% hydroxypropyl methylcellulose may have a viscosity of about 400 cps prior to instillation,yet unlike conventional high viscosity artificial tear formulations suchas Celluvisc® at about 400 cps, which may blur vision for 10-20 minutesor Liquigel® at about 100 cps, which causes similar but slightly reducedblurring causes only about 60 seconds of blur dissipating rapidly withan influx of tear secretion; where both a nonnewtonian reduction inviscosity at high shear (such as about 1/1000 sec during a blink, andaceclidine parasympathetic trigger of tear secretion as a sialogen maycontribute.

General miotic agents, such as pilocarpine, carbachol and phospholinediesterase, are capable of causing pupil miosis resulting in improvednear vision of presbyopic patients. However, there is an inversereduction in distance vision associated with these general miotic agentsfrom miosis at peak effect and accommodation that is not seen withaceclidine. The co-administration of a cycloplegic agent with aceclidinesurprisingly results in an attenuation of this reduction in distancevision.

Comfort, safety, and efficacy of a preferred embodiment of anophthalmological composition of the present invention results from thepresence of a nonionic surfactant, such as cyclodextrin alpha, beta, orgamma chains, preferably 2-hydroxypropyl beta-cyclodextrin (“HPβCD”),and, sulfobutyl ether derivative of β-cyclodextrin (Captisol®), apolyoxyl alkyl such as polyoxyl 40 stearate and polyoxyl 35 castor oil,or a poloxamer such as poloxamer 108 and poloxamer 407, a polysorbatesuch as polysorbate 80 or Brij® 35(Brij is a registered trademark ofUnigema Americas LLC); a viscosity enhancing agent, such ascarboxymethyl cellulose (“CMC”); a tonicity adjustor, such as sodiumchloride; a preservative, such as benzalkonium chloride and a pH fromabout 5.0 to about 8.0. Further, an increase in the concentration of thenonionic surfactant may result in reduced redness. Specifically,increasing polysorbate from 0.10% to 0.50-1.0% results in reducedredness. Further, increasing CMC or Carbopol® 940 from 0.50% to 1.5% w/v(preferably 1.40-1.43% w/v) results in enhanced near vision, bothquantitative improvement and duration improvement.

The viscosity of compositions of the present invention comprising aviscosity enhancer may be from about 1 to about 5,000 cps prior totopical instillation in the eye. As a result of the shear force appliedto the composition as it exits the device used for administration theviscosity is lowered to a range from about 1 to about 50 cps upontopical instillation, preferably from about 15 to about 35 cps.

The viscosity of compositions of the present invention comprising aviscosity agent may be about 25 cps or more at 25° C. and 0 shear, morepreferably from about 50 to about 10,000 cps, even more preferably fromabout 100 to about 5,000 cps and most preferably from about 150 to about450 cps prior to topical instillation in the eye. As a result of theshear force applied to the composition as it exits the device used foradministration the viscosity may be lowered to a range from about 1 toabout 25 cps at the high shear of blinking, and 50 cps to 200 cps at thelow shear between blinks, allowing greater drop retention with lessspillage and less nasolacrimal drainage and systemic absorption upontopical instillation.

Definitions

As used herein, the term “composition” is intended to encompass aproduct comprising the specified ingredients in the specified amounts,as well as any product which results, directly or indirectly, from acombination of the specified ingredients in the specified amounts.

The term “stabilizing”, as used herein, refers to any process whichfacilitates and/or enables an active agent to remain in solution. Theterm “stabilizing”, as used herein, also refers to any means or processwhich inhibits and/or reduces the tendency of a muscarinic agonist,including aceclidine, to degrade.

As used herein, all numerical values relating to amounts, weights, andthe like, that are defined as “about” each particular value is plus orminus 10%. For example, the phrase “about 5% w/v” is to be understood as“4.5% to 5.5% w/v.” Therefore, amounts within 10% of the claimed valueare encompassed by the scope of the claims.

As used herein “% w/v” refers to the percent weight of the totalcomposition.

As used herein the term “subject” refers but is not limited to a personor other animal.

The term muscarinic receptor agonist (“muscarinic agonist”) encompassesagonists that activate muscarinic acetylcholine receptors (“muscarinicreceptors”). Muscarinic receptors are divided into five subtypes namedM1-M5. Muscarinic agonists of the present invention include thosemuscarinic agonists that preferentially activate M1 and M3 receptorsover M2, M4 and M5 receptors (“M1/M3 agonists”). M1/M3 agonists includebut are not limited to aceclidine, xanomeline, talsaclidine,sabcomeline, cevimeline, alvameline, arecoline, milameline, SDZ-210-086,YM-796, RS-86, CDD-0102A(5-[3-ethyl-1,2,4-oxasdiazol-5-yl]-1,4,5,6-tetrahydropyrimidinehydrochloride), N-arylurea-substituted 3-morpholine arecolines,VUO255-035(N-[3-oxo-3-[4-(4-pyridinyl)-1-piperazinyl]propyl]-2,1,3-benzothiadiazole-4-sulfonamide),benzylquinolone carboxylic acid (BQCA), WAY-132983, AFB267B (NGX267),AC-42, AC-260584, chloropyrazines including but not limited to L-687,306, L-689-660, 77-LH-28-1, LY593039, and any quinuclidine ring with oneor more carbon substitutions particularly that include an ester, sulfur,or 5 or 6 carbon ring structure including with substituted nitrogen(s)and or oxygen(s), or any pharmaceutically acceptable salts, esters,analogues, prodrugs or derivatives thereof. A preferred M1/M3 agonist isaceclidine. In a preferred embodiment, muscarinic agonists of thepresent invention include those muscarinic agonist that preferentiallyactivate M1 and M3 over M2, M4, and M5; and even more preferablyactivate M1 over M3. In a more preferred embodiment muscarinic agonistof the present invention include those muscarinic agonists that onlyactivate M1.

The term “aceclidine” encompasses its salts, esters, analogues, prodrugsand derivatives including, but not limited to, aceclidine as a racemicmixture, aceclidine (R) enantiomer, aceclidine (S) enantiomer,aceclidine analogues, including, but not limited to, highly M1 selective1,2,5 thiadiazole substituted analogues like those disclosed in Ward. J.S. et al., 1,2,5-Thiadiazole analogues of aceclidine as potent mimuscarinic agonists, J Med Chem, 1998, January 29, 41(3), 379-392 andaceclidine prodrugs including but not limited to carbamate esters.

The term “selective α-2 adrenergic receptor agonists” or “α-2 agonist”encompasses all α-2 adrenergic receptor agonists which have a bindingaffinity of 900-fold or greater for α-2 over α-1 adrenergic receptors,or 300-fold or greater for α-2a or α-2b over α-1 adrenergic receptors.

The term also encompasses pharmaceutically acceptable salts, esters,prodrugs, and other derivatives of selective α-2 adrenergic receptoragonists.

The term “low concentrations” or “low-dose” of alpha-2 adrenergicreceptor agonists refers to concentrations from between about 0.0001% toabout 0.065% w/v; more preferably, from about 0.001% to about 0.035%w/v; even more preferably, from about 0.01% to about 0.035% w/v; andeven more preferably, from about 0.03% to about 0.035% w/v.

The term “brimonidine” encompasses, without limitation, brimonidinesalts and other derivatives, and specifically includes, but is notlimited to, brimonidine tartrate,5-bromo-6-(2-imidazolin-2-ylamino)quinoxaline D-tartrate, and Alphagan®.

The terms “treating” and “treatment” refer to reversing, alleviating,inhibiting, or slowing the progress of the disease, disorder, orcondition to which such terms apply, or one or more symptoms of suchdisease, disorder, or condition.

The term “pharmaceutically acceptable” describes a material that is notbiologically or otherwise undesirable (i.e. without causing anunacceptable level of undesirable biological effects or interacting in adeleterious manner).

As used herein, the term “pharmaceutically effective amount” refers toan amount sufficient to affect a desired biological effect, such as abeneficial result, including, without limitation, prevention,diminution, amelioration or elimination of signs or symptoms of adisease or disorder. Thus, the total amount of each active component ofthe pharmaceutical composition or method is sufficient to show ameaningful subject benefit. Thus, a “pharmaceutically effective amount”will depend upon the context in which it is being administered. Apharmaceutically effective amount may be administered in one or moreprophylactic or therapeutic administrations.

The term “prodrugs” refers to compounds, including, but not limited to,monomers and dimers of the compounds of the invention, which havecleavable groups and become, under physiological conditions, compoundswhich are pharmaceutically active in vivo.

As used herein “salts” refers to those salts which retain the biologicaleffectiveness and properties of the parent compounds and which are notbiologically or otherwise harmful at the dosage administered. Salts ofthe compounds of the present inventions may be prepared from inorganicor organic acids or bases.

The term “higher order aberrations” refers to aberrations in the visualfield selected from starbursts, halos (spherical aberration), doublevision, multiple images, smeared vision, coma and trefoil.

The term “cold chain” refers to storage at temperatures from about 2 toabout 8° C. from manufacture to immediately prior to administration.

The compounds of the present invention can be used in the form ofpharmaceutically acceptable salts derived from inorganic or organicacids or bases. The phrase “pharmaceutically acceptable salt” meansthose salts which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of humans and lower animalswithout undue toxicity, irritation, allergic response and the like andare commensurate with a reasonable benefit/risk ratio. Pharmaceuticallyacceptable salts are well-known in the art. For example, S. M. Berge etal. describe pharmaceutically acceptable salts in detail in J.Pharmaceutical Sciences, 1977, 66: 1 et seq.

The salts can be prepared in situ during the final isolation andpurification of the compounds of the invention or separately by reactinga free base function with a suitable organic acid. Representative acidaddition salts include, but are not limited to acetate, adipate,alginate, citrate, aspartate, benzoate, benzenesulfonate, bisulfate,butyrate, camphorate, camphorsulfonate, digluconate, glycerophosphate,hemisulfate, heptanoate, hexanoate, fumarate, hydrochloride,hydrobromide, hydroiodide, 2-hydroxyethansulfonate (isothionate),lactate, maleate, methanesulfonate, nicotinate, 2-naphthalenesulfonate,oxalate, palmitoate, pectinate, persulfate, 3-phenylpropionate, picrate,pivalate, propionate, succinate, tartrate, thiocyanate, phosphate,glutamate, bicarbonate, p-toluenesulfonate and undecanoate. Also, thebasic nitrogen-containing groups can be quaternized with such agents aslower alkyl halides such as methyl, ethyl, propyl, and butyl chlorides,bromides and iodides; dialkyl sulfates like dimethyl, diethyl, dibutyland diamyl sulfates; long chain halides such as decyl, lauryl, myristyland stearyl chlorides, bromides and iodides; arylalkyl halides likebenzyl and phenethyl bromides and others. Water or oil-soluble ordispersible products are thereby obtained. Examples of acids which canbe employed to form pharmaceutically acceptable acid addition saltsinclude such inorganic acids as hydrochloric acid, hydrobromic acid,hyaluronic acid, malic acid, sulphuric acid and phosphoric acid and suchorganic acids as oxalic acid, malic acid, maleic acid, methaneosulfonicacid, succinic acid and citric acid.

Basic addition salts can be prepared in situ during the final isolationand purification of compounds of this invention by reacting a carboxylicacid-containing moiety with a suitable base such as the hydroxide,carbonate or bicarbonate of a pharmaceutically acceptable metal cationor with ammonia or an organic primary, secondary or tertiary amine.Pharmaceutically acceptable salts include, but are not limited to,cations based on alkali metals or alkaline earth metals such as lithium,sodium, potassium, calcium, magnesium and aluminum salts and the likeand nontoxic quaternary ammonia and amine cations including ammonium,tetramethylammonium, tetraethylammonium, methylammonium,dimethylammonium, trimethylammonium, triethylammonium, diethylammonium,and ethylammonium among others. Other representative organic aminesuseful for the formation of base addition salts include ethylenediamine,ethanolamine, diethanolamine, piperidine, piperazine and the like.

The term “ester” as used herein is represented by the formula —OC(O)A¹or —C(O)OA¹, where A¹ can be alkyl, cycloalkyl, alkenyl, cycloalkenyl,alkynyl, cycloalkynyl, aryl, a heteroaryl group or other suitablesubstituent.

Compositions of the Invention

In one embodiment, the present invention is directed to anophthalmological composition comprising aceclidine. In a preferredembodiment, aceclidine is at a concentration from about 0.25% to about2.0% w/v, more preferably from about 0.75% to about 2.5% w/v, still morepreferably from about 1.65% to about 1.85% w/v, and most preferablyabout 1.75% w/v. In a preferred embodiment, aceclidine is at aconcentration from about 0.25% to about 2.0% w/v, more preferably fromabout 0.50% to about 1.90% w/v, still more preferably from about 1.25%to about 1.85% w/v, and most preferably from about 1.35% to about 1.65%w/v. As aceclidine is a tertiary amine with asymmetry, both a + and −optical isomer exists (where in some studies (+) is more potent and inothers it is felt (−) may be more potent). For the above concentrationspolarimetry demonstrated an exactly equal ratio of (+) and (−) isomerfor these concentrations. Altering this ratio could therefore alter thisconcentration range proportional to a change in ratio.

The present invention is further directed to an ophthalmologicalcomposition comprising a muscarinic agonist, preferably a nonionicsurfactant above its critical micellar concentration for thecomposition, and a viscosity enhancing agent; or alternatively anin-situ gelling agent. In preferred embodiments the initial viscosity ofthe composition on topical application is above 20 cps, preferably above50 cps, and more preferably at about 65 cps or more at 25° C. and 0shear.

Cryoprotectants are compounds that either prevent freezing or preventdamage to compounds during freezing. As used herein, the term“cryoprotectant” or “cryoprotectants” include lyoprotectants.Cryoprotectants suitable for use in the subject invention include, butare not limited to, a polyol, a sugar, an alcohol, a lower alkanol, alipophilic solvent, a hydrophilic solvent, a bulking agent, asolubilizer, a surfactant, an antioxidant, a cyclodextrin, amaltodextrin, colloidal silicon dioxide, polyvinyl alcohol, glycine,2-methyl-2,4-pentanediol, cellobiose, gelatin, polyethylene glycol(PEG), dimethyl sulfoxide (DMSO), formamide, antifreeze protein 752 or acombination thereof.

As used herein the term “polyol” refers to compounds with multiplehydroxyl functional groups available for organic reactions such asmonomeric polyols such as glycerin, pentaerythritol, ethylene glycol andsucrose. Further, polyols may refer to polymeric polyols includingglycerin, pentaerythritol, ethylene glycol and sucrose reacted withpropylene oxide or ethylene oxide. In a preferred embodiment, polyolsare selected from the group consisting of mannitol, glycerol,erythritol, lactitol, xylitol, sorbitol, isosorbide, ethylene glycol,propylene glycol, maltitol, threitol, arabitol and ribitol. In a morepreferred embodiment, the polyol is mannitol.

Sugars suitable for use in the present invention as cryoprotectantsinclude, but are not limited to, glucose, sucrose, trehalose, lactose,maltose, fructose and dextran.

In another preferred embodiment, alcohols include, but are not limitedto, methanol.

In one embodiment, the present invention individually excludes eachcryoprotectant from the definition of cryoprotectant.

Cryoprotectants may be at present in compositions of the presentinvention at a concentration from about 0.1% to about 99% w/v,preferably from about 1% to about 50% w/v, more preferably from about 1%to about 10% w/v.

As used herein “lower alkanols” include C1-C6 alkanols. Lower alkanols,suitable for use in the present invention include, but are not limitedto, amyl alcohol, butanol, sec-butanol, t-butyl alcohol, n-butylalcohol, ethanol, isobutanol, methanol. isopropanol and propanol.

Bulking agents suitable for use in the present invention include, butare not limited to, saccharide, polyvinylpyrrolidone, cyclodextrin andtrehalose.

Solubilizers suitable for use in the present invention include, but arenot limited to, cyclic amide, gentisic acid and cyclodextrins.

In a preferred embodiment, surfactants suitable for use in the presentinvention include, but are not limited to, nonionic surfactants, morepreferably surfactants with a hydrophilic-lipophilic balance (“HLB”)value of 1 to 18.

In a preferred embodiment, antioxidants suitable for use in the presentinvention include, but are not limited to, bisulfite, ascorbic acid,disodium- or tetrasodium ethylenediaminetetraacetic acid, citrate,butylated hydroxyanisole (“BHA”), butylated hydroxytoluene (“BHT”), asulfoxylate, propyl gallate, an amino acid containing a thio group, anda thiol.

Nonionic surfactants suitable for the present invention includecyclodextrins, polyoxyl alkyls, poloxamers or combinations thereof, andmay include in addition combinations with other nonionic surfactantssuch as polysorbates. Nonionic surfactants suitable for the presentinvention include a polysorbate, tyloxapol, a poloxamer, a cyclodextrin,vitamin E TPGS, a polyoxyl castor oil, a polyoxyl stearate, polyethyleneglycol, a polyoxyethylene glycol alkyl ether and2-[[10,13-dimethyl-17-(6-methylheptan-2-yl)-2,3,4,7,8,9,11,12,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-3-yl]oxy]ethanol.Preferred embodiments include Poloxamer 80, Poloxamer 188, Poloxamer407, Polysorbate 20, Polysorbate 80, ionically charged (e.g. anionic)beta-cyclodextrins with or without a butyrated salt (Captisol®)2-hydroxypropyl beta cyclodextrin (“HPOCD”), alpha cyclodextrins, gammacyclodextrins, Polyoxyl 35 castor oil, and Polyoxyl 40 hydrogenatedcastor oil or combinations thereof. Further, substitution of othernonionic surfactants compatible with ophthalmological use allows forsimilar formulation advantages, which may include but is not limited toone or more of a nonionizing surfactant such as poloxamer, poloxamer103, poloxamer 123, and poloxamer 124, poloxamer 407, poloxamer 188, andpoloxamer 338, any poloxamer analogue or derivative, polysorbate,polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, anypolysorbate analogue or derivative, cyclodextrin,hydroxypropyl-β-cyclodextrin, hydroxypropyl-γ-cyclodextrin, randomlymethylated β-cyclodextrin, β-cyclodextrin sulfobutyl ether,γ-cyclodextrin sulfobutyl ether or glucosyl-β-cyclodextrin, anycyclodextrin analogue or derivative, polyoxyethylene, polyoxypropyleneglycol, an polysorbate analogue or derivative, polyoxyethylenehydrogenated castor oil 60, polyoxyethylene (200), polyoxypropyleneglycol (70), polyoxyethylene hydrogenated castor oil, polyoxyethylenehydrogenated castor oil 60, polyoxyl, polyoxyl stearate, nonoxynol,octyphenol ethoxylates, nonyl phenol ethoxylates, capryols, lauroglycol,polyethylene glycol (“PEG”), Brij® 35, 78, 98, 700 (polyoxyethyleneglycol alkyl ethers), glyceryl laurate, lauryl glucoside, decylglucoside, or cetyl alcohol; or zwitterion surfactants such as palmitoylcarnitine, cocamide DEA, cocamide DEA derivatives cocamidopropylbetaine, or trimethyl glycine betaine, N-2(2-acetamido)-2-aminoethanesulfonic acid (ACES), N-2-acetamido iminodiacetic acid (ADA),N,N-bis(2-hydroxyethyl)-2-aminoethane sulfonic acid (BES),2-[Bis-(2-hydroxyethyl)-amino]-2-hydroxymethyl-propane-1,3-diol(Bis-Tris), 3-cyclohexylamino-1-propane sulfonic acid (CAPS),2-cyclohexylamino-1-ethane sulfonic acid (CHES),N,N-bis(2-hydroxyethyl)-3-amino-2-hydroxypropane sulfonic acid (DIPSO),4-(2-hydroxyethyl)-1-piperazine propane sulfonic acid (EPPS),N-2-hydroxyethylpiperazine-N′-2-ethane sulfonic acid (HEPES),2-(N-morpholino)-ethane sulfonic acid (MES), 4-(N-morpholino)-butanesulfonic acid (MOBS), 2-(N-morpholino)-propane sulfonic acid (MOPS),3-morpholino-2-hydroxypropanesulfonic acid (MOPSO),1,4-piperazine-bis-(ethane sulfonic acid) (PIPES),piperazine-N,N′-bis(2-hydroxypropane sulfonic acid) (POPSO),N-tris(hydroxymethyl)methyl-2-aminopropane sulfonic acid (TAPS),N-[tris(hydroxymethyl)methyl]-3-amino-2-hydroxypropane sulfonic acid(TAPSO), N-tris(hydroxymethyl) methyl-2-aminoethane sulfonic acid (TES),2-Amino-2-hydroxymethyl-propane-1,3-diol (Tris), tyloxapol, Solulan™C-24(2-[[10,13-dimethyl-17-(6-methylheptan-2-yl)-2,3,4,7,8,9,11,12,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-3-yl]oxy]ethanol)and Span® 20-80 (sorbitan monolaurate, sorbitan monopalmitate, sorbitanmonostearate, and sorbitan monooleate). In certain embodiments theaddition of an anionic surfactant such as sodium lauryl sulfate and orsodium ester lauryl sulfate may be preferred. In other embodiments theaddition of polysorbate 80 is preferred. In addition to the abovenonionic surfactants any nonionic surfactant is suitable for use in thepresent invention as long as the concentration of the nonionicsurfactant is such that it is above the critical micellar concentrationfor that non-ionic surfactant. Preferably, the nonionic surfactants usedin the present invention achieve submicron diameter micelles, morepreferably less than 200 nanometers and more preferably less than 150nanometers in diameter.

Ophthalmological in situ gels which may be substituted for or added inaddition to one or more nonionic surfactants include but are not limitedto gelatin, carbomers of various molecular weights including carbomer934 P and 974 P, xanthan gums, alginic acid (alginate), guar gums,locust bean gum, chitosan, pectins and other gelling agents well knownto experts in the art.

In preferred embodiments the nonionic surfactant is polyoxyl 40 stearateat a concentration from about 1 to about 15% w/v, more preferably atabout 5.5% w/v.

In other preferred embodiments, the nonionic surfactant is polysorbate80 at a concentration from about 0.5% to about 10% w/v, more preferablyfrom about 1% to about 7% w/v and even more preferably from about 2% toabout 5% w/v, yet more preferably from about 2.5% to about 4% w/v andmost preferably at about 2.5% or 2.75% or 3% or 4% or 5% w/v.

Viscosity agents suitable for the present invention include, but are notlimited to gums such as guar gum, hydroxypropyl-guar (“hp-guar”), andxanthan gum, alginate, chitosan, gelrite, hyaluronic acid, dextran,Carbopol® (polyacrylic acid or carbomer) including Carbopol® 900 seriesincluding Carbopol® 940 (carbomer 940), Carbopol® 910 (carbomer 910) andCarbopol® 934 (carbomer 934), cellulose derivatives such as highmolecular weight carboxymethyl cellulose (“CMC”), methylcellulose,methyl cellulose 4000, hydroxymethyl cellulose, hydroxypropyl cellulose,hydroxypropylmethyl cellulose (hypromellose), hydroxyl propyl methylcellulose 2906, carboxypropylmethyl cellulose, hydroxypropylethylcellulose, and hydroxyethyl cellulose, dextran, polyethylene glycol,polyvinyl alcohol, polyvinyl pyrrolidone, gellan, carrageenan, alginicacid, carboxyvinyl polymer or combinations thereof.

In a preferred embodiment the viscosity agent will provide a viscosityof the total composition from about 50 to about 10,000 cps, from about100 to about 5,000 cps or from 150 to about 450 cps at 25° C. and 0shear.

In another preferred embodiment the viscosity agent will have anequilibration viscosity less than 100 cps, preferably from about 15 toabout 35 cps, and most preferably at about 30 cps. In a preferredembodiment the viscosity agent is Carbopol® 940 (carbomer 940) at aconcentration from about 0.05% to about 1.5% w/v, preferably from about0.09% to about 1.0% w/v, more preferably at 0.09%, 0.25%, 0.5%, 0.75%,0.9% or 1.0% w/v. In certain combinations it has been surprisinglydiscovered nonionic surfactant/viscosity combinations may result inphase separation over time with precipitate formation. In suchsituations, particularly for polyoxyls, in a preferred embodimentpolyoxyl 40 stearate, and cellulose derivatives, particularlyhydroxypropylmethyl cellulose, use of a nonpolysaccharide derivative forviscosity enhancement, such as polyacrylic acid derivatives (carbomers,carbomer 934 or 940 in preferred embodiments) may prevent suchseparation; or alternatively use of a non polyoxyl nonionic surfactant,such as polysorbate 80 with either a cellulose derivative ornoncellulose derivative viscosity agent may be substituted.

In another preferred embodiment, the viscosity agent is carboxymethylcellulose at a concentration from about 0.1% to about 2% w/v, morepreferably from 0.25% to about 1.3% w/v, when at a viscosity of 3,500cps at 25° C. in a 2% solution.

In another preferred embodiment, the viscosity agent ishydroxypropylmethyl cellulose at a concentration from about 0.5% toabout 1.75%, and more preferably about 0.75% or 1.5%, still morepreferably from about 1.0% to about 1.5%, and most preferably at about1.25%.

Not wishing to be held to particularly theory, it appears thequinuclidine nucleus of the heterocyclic nitrogen on aceclidine is soelectron rich it easily attacks surrounding compounds as well as itself.

It is a discovery of the present invention that several modificationsmay singly or in combination be used to enhance cold chain stabilitystorage, including in addition to in a preferred embodiment aceclidine1.40%-1.75%, tropicamide 0.025%-0.10% and optionally a nonioinicsurfactant such as polyoxyl 40 stearate 0.5%-10%, preferably 5.5% one ormore of (See Table 3):

-   -   Acidic pH, preferably less than 5.5, preferably less than 5.0        and most preferably at a pH of about 4.75;    -   Viscosity agent, preferably at 25° C. viscosity of about 15-50        cps, and more preferably 20-45 cps, where a preferred embodiment        is carbomer 940 0.09%-1.5%;    -   Addition of a cryoprotectant, in a preferred embodiment a        polyol, preferably Mannitol 2.5%-4.0%;    -   Addition of a buffer, where acetate or phosphate buffers are        preferred, 2-100 mmole range with 3-5 mmole is preferred; and    -   Addition of a preservative, where BAK 0.015% is preferred.

The selective α-2 agonist may be included within the composition of thepresent invention or applied topically preferably just minutes before orless preferably just minutes afterward if additional means to reducenasal congestion or redness is desired for sensitive subjects. Selectiveα-2 agonists suitable for the present invention have minimal α-1 agonistactivity at low concentrations. For example, for brimonidine orfadolmidine, 1% to 2% w/v is considered extremely high, 0.5% to 1.0% w/vstill highly inductive of α-1 receptors and toxic for purposes of thepresent invention. Further, 0.10% to 0.5% w/v is still too high and even0.070% to 0.10% w/v is associated with a higher than preferred incidenceof rebound hyperemia (however, for dexmedetomidine, its greaterlipophilicity and intraocular penetration reduces rebound risk in thisrange). Only 0.065% w/v or below is potentially acceptable, where formost α-2 agonists, depending on degree of selectivity 0.050% w/v or evenmore preferably 0.035% w/v or less is desired. On the other hand, somedegree of useful activity may occur at one or more orders of magnitudefurther reduction of concentration. The preferred embodiments,brimonidine, fadolmidine and guanfacine, of the present inventionpreferentially stimulate α-2 adrenergic receptors, and even morepreferably α-2b adrenergic receptors so that α-1 adrenergic receptorsare not stimulated sufficiently enough to cause excessive large vesselarteriolar constriction and vasoconstrictive ischemia. In addition, ithas been discovered that preventing or reducing redness for drugs thatotherwise directly induce redness, such as the acetylcholine agonist,aceclidine, enhances compliance for sensitive subjects that may haveinduced redness or nasal congestion even with formulations of thepresent invention that do not include an α-2 agonist. However, becauseα-2 agonists are shifted to their ionized equilibrium an acidic pH issomewhat offset by the fact such agonists exert greater affect atneutral or alkaline pH.

Therefore, each α-2 agonist has a preferred pH range depending on itslipophilicity and pKa value when added to the inventive compositionswith aceclidine. For the present invention while pH range of 5.0 to 8.0is tolerated, preferred embodiments are at pH 5.5 to 7.5 and morepreferably 6.5 to 7.0. Further, it has been discovered thatcyclodextrins and/or polyoxyl 40 stearate as a nonionic surfactantcomponent or as the sole nonionic surfactant, result in a greaterwhitening effect when the α-2 agonist is included in the compositionrather than poloxamer 407. The α-2 agonist may optionally be appliedseparately or in certain preferred embodiments with formulations of thepresent invention that do not include an α-2 agonist, such as thoseformulas with polyoxyl 40 stearate 5.5% w/v as the non-ionic surfactant,although the α-2 agonist is not required except for occasional sensitivesubjects. Fadolmidine represents the α-2 agonist with highesthydrophilicity and therefore high surface retention for the presentinvention. Guanfacine is also highly selective and hydrophilic.Brimonidine is highly selective with moderate lipophilicity. Finally,dexmedetomidine has high selectivity with high lipophilicity that may beused with less efficacy for reducing redness for the purposes of thepresent invention (although possibly inducing fatigue as a side effectin some patients). In a preferred embodiment using polyoxyl 40 stearate5.5% w/v; CMC 0.80% w/v; NaCl 0.037% w/v; ethylenediaminetetraaceticacid (“EDTA”) 0.015% w/v, borate buffer 5 mM and BAK 0.007% w/v resultsin redness of about 1.0 to 1.5 out of 4 which is transient lasting aboutten minutes, and by 30 minutes returns to about baseline.

In one embodiment, the selective α-2 adrenergic receptor agonist is acompound which has binding affinity of about 900-fold or greater, evenmore preferably about 1000-fold or greater, and most preferably, about1500-fold or greater.

The selective α-2 adrenergic receptor agonist may be present at aconcentration from between about 0.0001% to about 0.065% w/v; morepreferably, from about 0.001% to about 0.035% w/v; even more preferably,from about 0.01% to about 0.035% w/v; and even more preferably, fromabout 0.020% to about 0.035% w/v.

In one embodiment, the selective α-2 adrenergic receptor is selectedfrom the group consisting of brimonidine, guanfacine, fadolmidine,dexmedetomidine,(+)-(S)-4-[1-(2,3-dimethyl-phenyl)-ethyl]-1,3-dihydro-imidazole-2-thione,1-[(imidazolidin-2-yl)imino]indazole, and mixtures of these compounds.Analogues of these compounds that function as highly selective α-2agonists may also be used in compositions and methods of the presentinvention.

In a more preferred embodiment, the selective α-2 agonist is selectedfrom the group consisting of fadolmidine, guanfacine and brimonidine. Ina yet more preferred embodiment the selective α-2 agonist is brimonidinein the form of a salt at a concentration of 0.025% to 0.065% w/v, morepreferably from 0.03% to 0.035% w/v. In a preferred embodiment, the saltis a tartrate salt.

In another yet more preferred embodiment, the selective α-2 agonist isfadolmidine at a concentration from about 0.005% to about 0.05% w/v,more preferably from 0.02% to about 0.035% w/v in the form of ahydrochloride (“HCl”) salt.

In another yet more preferred embodiment, the selective α-2 agonist isguanfacine at a concentration from about 0.005% to about 0.05% w/v, morepreferably from 0.02% to about 0.035% w/v in the form of an HCl salt.

In another yet more preferred embodiment, the selective α-2 agonist isdexmedetomidine at a concentration from about 0.005% to about 0.05% w/v,more preferably from 0.04% to about 0.05% w/v in the form of an HClsalt.

In another preferred embodiment a pH less than physiologic pH is foundto enhance the whitening effect for brimonidine, preferably pH 4.5 to6.5, and more preferably pH 5.5 to 6.0.

However, redness reduction is achieved at all pHs, and enhancement ofaceclidine absorption occurs at alkaline pH, such that more effectoccurs from a given concentration, and therefore while effective at pHranges from 4.5 to 8.0, pH range of 6.5 to 7.5 is preferred for thepresent invention, and 7.0 to 7.5 most preferred.

The present invention is further directed to an ophthalmologicalcomposition further comprising a cycloplegic agent. It is a surprisingand totally unexpected discovery of the present invention that certaincycloplegic agents can be combined with miotic agents, particularly forthe present invention, aceclidine, without reducing miotic onset,magnitude, or duration; and further blunt the normally attendant spikein miotic effect coinciding with time of peak absorption in aqueousformulations to provide a constant miosis versus time after onset from15 to 30 minutes to 6 to 10 hours depending on the desired formulation.The addition of the cycloplegic agent also reduces any residualassociated discomfort that may otherwise occur soon after topicalinstillation, which presumably is a result of ciliary spasms orexcessive pupillary miosis.

Cycloplegic agents suitable for the present invention include, but arenot limited to, atropine, Cyclogyl® (cyclopentolate hydrochloride),hyoscine, pirenzepine, tropicamide, atropine,4-diphenylacetoxy-N-methylpiperidine methobromide (4-DAMP), AF-DX 384,methoctramine, tripitramine, darifenacin, solifenacin (Vesicare),tolterodine, oxybutynin, ipratropium, oxitropium, tiotropium (Spriva),and otenzepad (a.k.a. AF-DX 116 or11-{[2-(diethylamino)methyl]-1-piperidinyl}acetyl]-5,11-dihydro-6H-pyrido[2,3b][1,4]benzodiazepine-6-one).In a preferred embodiment the cycloplegic agent is tropicamide at aconcentration from about 0.004% to about 0.025% w/v, more preferablyfrom about 0.005% to about 0.015% w/v and still more preferably fromabout 0.005% to about 0.011% w/v, from about 0.005% to about 0.007% w/vand from about 0.005% to about 0.006% w/v. In another preferredembodiment the cycloplegic agent is a mixture of tropicamide at aconcentration from about 0.04% to about 0.07% w/v or pirenzepine orotenzepad at a concentration from about 0.002% to about 0.05% w/v.

In a preferred embodiment, tropicamide 0.01% w/v was found to slightlyreduce brow ache, 0.030% w/v to further reduce brow ache and from 0.04%to about 0.07% w/v to completely eliminate brow ache without reductionof the average pupillary miosis diameter over duration of effect.Tropicamide in preferred embodiments has demonstrated completelyunexpected sensitivity of effect, where at about 0.04% w/v unexpectedlyand very effectively reduces or eliminates brow ache and ciliary spasmpain, becoming very noticeably further reduced at 0.042% w/v and absentat 0.044% w/v in a preferred embodiment with no cycloplegia (surprisingdue to its common use as a pupil dilating agent). Yet, tropicamide didnot reduce the mean degree of pupil miosis, the time of onset of pupilmiosis or the subsequent visual benefits. On the contrary, tropicamideblunted the peak miosis seen in aqueous formulations to create a smoothconsistent miotic effect over time. It allowed modulation of peak pupilmiosis to achieve a more even effect over time with no dilation as hasbeen found with its prior use. Specifically, tropicamide is useful toprevent transient constriction below 1.50 mm at 30 to 60 minutesfollowing aceclidine in some embodiments and to reduce transientexcessive and undesirable dimming of vision that may otherwise occur atpeak onset of about 30 minutes. As an example, an ophthalmologicalcomposition comprising 1.53% w/v aceclidine, 5% w/v HPβCD, 0.75% w/vCMC, 0.25% w/v NaCl, 0.01% w/v BAK and a phosphate buffer at pH 7.0; or1.45% w/v aceclidine; 5.5% w/v polyoxyl 40 stearate; 0.80% w/v CMC;0.037% w/v NaCl; 0.015% w/v EDTA; 0.007% w/v BAK and 5 mM phosphatebuffer at a pH 7.0; was varied from 0.040% w/v tropicamide, wheremoderate dimming was noted, to 0.044% w/v tropicamide where dimmingbecame almost undetectable other than in extremely dim light conditions.This additional pupil size modulation with a cycloplegic agent allowsaceclidine concentrations sufficient for prolonged effect while bluntingthe attendant peak excessive constriction that is undesirable as well asany uncomfortable brow ache. Surprisingly and due to its short-actingnature, tropicamide achieves this blunting effect without causingmydriasis. Further, in a preferred embodiment, tropicamide 0.014% w/vwas found to reduce brow ache, 0.021% w/v to further reduce brow acheand from 0.028% to 0.060% w/v and in some embodiments up to 0.09% w/v tocompletely eliminate brow ache without cycloplegia (i.e. paralysis ofciliary muscle of the eye).

It has been found for a racemic 50:50 mixture of (+) and (−) aceclidineoptical isomers (where in some studies (+) is more potent and in othersit is felt (−) may be more potent) tropicamide effects may varydepending on the ratio of aceclidine to tropicamide. For example, in anophthalmological composition of the present invention comprising 1.55%w/v aceclidine, 5.5% w/v HPβCD or in a preferred embodiment polyoxyl 40stearate, 0.75% w/v CMC (1%=2,500 centipoise), 0.25% w/v NaCl, and 0.01%w/v BAK and at pH 7.5, 0.042% w/v tropicamide can be differentiated fromeven 0.035% w/v, with the former demonstrating normal indoor nightvision and the latter slight dimming that becomes more noticeable atstill lower concentrations. At higher concentrations, such as from about0.075% to about 0.090% w/v tropicamide, loss of optimal range pupilconstriction 1.50 mm to 1.80 mm range begins, and frank mydriasis athigher concentrations begins to occur. As isomer ratio may alter theeffective concentration, this must be factored into the clinicalefficacy anticipated using aceclidine; for preferred embodiments of thepresent invention a polarimeter was used to determine an exact 50:50isomer ratio was used (personal communication Toronto ResearchChemicals).

FIG. 1 shows the effect of a miotic agent with or without a cycloplegicagent and with or without a carrier. Subject is an emmetrope over theage of 45 with a baseline near vision of 20.100 and baseline distancevision of 20.20. Topical administration to the eye of 1% w/v pilocarpinein saline solution results in an improvement of near vision to 20.40(8a), however this improvement comes at the expense of a reduction indistance vision to 20.100 (8b). The addition of 0.015% w/v tropicamideresults in an improvement of near vision to 20.25 (9a) and a lesseningof the reduction of distance vision to 20.55 (9b), though in certaininstances with some induced irregular astigmatism (mildly blotched areasin reading field of vision). Topical administration of 1.55% w/vaceclidine in saline solution results in an improvement of near visionto 20.40 for an extended time period of 6 hrs (10a) without any effecton the baseline distance vision (10b). 10c and 10d show the effects ofadministering aceclidine in a carrier composed of 5.5% w/v2-hydroxypropyl beta cyclodextrin, 0.75% w/v CMC (1%=2,500 centipoise),0.25% w/v NaCl, and 0.01% w/v BAK. As seen in 10c the carrier increasesthe beneficial effect of aceclidine resulting in better than 20.20 nearvision. As seen in 10d a similar increase in distance vision occurs. 10eand 10f show the effects of adding 0.042% w/v tropicamide to theaceclidine in the carrier. As seen in 10e near vision is improved to20.15 with a quicker onset of maximum visual acuity. As seen in 10f asimilar improvement is seen in distance vision. Taken together, FIG. 1shows that aceclidine is capable of temporarily correcting near visionin a presbyopic subject without affecting the baseline distance vision.Similar results can be achieved with a different miotic agent,pilocarpine, with the addition of a cycloplegic agent such astropicamide. A proper drug carrier can also have a beneficial effect.

The present invention is further directed to an ophthalmologicalcomposition further comprising a tonicity adjustor and a preservative.

A tonicity adjustor can be, without limitation, a salt such as sodiumchloride (“NaCl”), potassium chloride, mannitol or glycerin, or anotherpharmaceutically or ophthalmologically acceptable tonicity adjustor.

Preservatives that can be used with the present invention include, butare not limited to, benzalkonium chloride (“BAK”), sorbic acid,oxychloro complex, citric acid, chlorobutanol, thimerosal,phenylmercuric acetate, disodium ethylenediaminetetraacetic acid,phenylmercuric nitrate, perborate or benzyl alcohol. In a preferredembodiment the preservative is BAK, sorbic acid, oxychloro complex or acombination thereof. In a yet more preferred embodiment BAK is at aconcentration of about 0.001% to about 1.0% w/v, more preferably at aconcentration of about 0.007%, 0.01% or 0.02% w/v. In another preferredembodiment the preservative is perborate at a concentration of 0.01% toabout 1.0% w/v, more preferably at a concentration of about 0.02% w/v.

Various buffers and means for adjusting pH can be used to prepareophthalmological compositions of the invention. Such buffers include,but are not limited to, acetate buffers, citrate buffers, phosphatebuffers and borate buffers. It is understood that acids or bases can beused to adjust the pH of the composition as needed, preferably of 1 to10 mM concentration, and more preferably about 3 mM or 5 mM. In apreferred embodiment the pH is from about 4.0 to about 8.0, in a morepreferred embodiment the pH is from about 5.0 to about 7.0.

The present invention is further directed to an ophthalmologicalcomposition further comprising an antioxidant. Antioxidants that can beused with the present invention include but are not limited to disodiumethylenediaminetetraacetic acid or disodium edetate dihydrate at aconcentration from about 0.005% to about 0.50% w/v, citrate at aconcentration from about 0.01% to about 0.3% w/w, dicalciumdiethylenetriamine pentaacetic acid (“Ca2DTPA”) at a concentration fromabout 0.001% to about 0.2% w/v, preferably about 0.01% w/v Ca2DTPA whichcan be formulated by adding 0.0084% w/v Ca(OH)₂ and 0.0032% w/v penteticacid to the formulation and mixing slowly. Further combinations ofantioxidants can be used. Other antioxidants that can be used with thepresent invention include those well known to experts in the art such asethylenediaminetetraacetic acid at a concentration from about 0.0001% toabout 0.015% w/v.

It is a surprising and unexpected discovery that topical formulations ofthe present invention, particularly one of the preferred embodimentscomprising aceclidine 1.35% to 1.55% w/v; 5.5% w/v polyoxyl 40 stearate;0.80% w/v CMC; 0.037% w/v NaCl; 0.015% w/v EDTA; 0.007% w/v BAK; and 5mM phosphate buffer at pH 7.0 result in considerably prolonged contactlens wear and comfort after a single topical instillation daily. Thesingle daily use of the preferred embodiments allowed a subject with dryeye to sleep in his lenses for one-week periods where previously evenafter a single night vision would be blurred and contact lenses coatedwith film requiring removal and cleaning or replacement (see Example 7).

In preferred embodiments, an ophthalmological composition of the presentinvention comprises aceclidine, a cryoprotectant, optionally acycloplegic agent, a nonionic surfactant at a concentration from about1% to about 5% w/v and a viscosity agent at a concentration of about0.75% to about 1.6% w/v, preferably about 1.25% to about 1.5% w/v.

The following representative embodiments are provided solely forillustrative purposes and are not meant to limit the invention in anyway.

REPRESENTATIVE EMBODIMENTS

In one embodiment, the ophthalmological composition comprises:

-   -   aceclidine at a concentration of about 1.75% w/v; and    -   mannitol at a concentration of about 2.5% w/v.

In another embodiment, the ophthalmological composition comprises:

-   -   aceclidine at a concentration of about 1.75% w/v;    -   mannitol at a concentration of about 2.5% w/v; and tropicamide        at a concentration of about 0.02% w/v.

In another embodiment, the ophthalmological composition comprises:

-   -   aceclidine at a concentration of about 1.75% w/v;    -   mannitol at a concentration of about 2.5% w/v;    -   polysorbate 80 at a concentration of about 5.0% w/v;    -   carboxymethyl cellulose at a concentration of about 1.4% w/v;    -   BAK at a concentration of about 0.015% w/v; and    -   phosphate buffer at a concentration of about 3 mM,    -   wherein the pH is about 5.

In another embodiment, the ophthalmological composition comprises:

-   -   aceclidine at a concentration of about 1.75% w/v;    -   mannitol at a concentration of about 2.5% w/v;    -   polysorbate 80 at a concentration of about 0.5% w/v;    -   NaCl at a concentration from about 0.10% to about 0.50% w/v;    -   Carbopol® 940 at a concentration of about 0.95% w/v;    -   BAK at a concentration of about 0.01% w/v; and    -   phosphate buffer at a concentration of about 3 mM,    -   wherein the pH is about 5.

In another embodiment, the ophthalmological composition comprises:

-   -   aceclidine at a concentration of about 1.75% w/v;    -   mannitol at a concentration of about 2.5% w/v;    -   polysorbate 80 at a concentration of about 2.0% w/v;    -   NaCl at a concentration of about 0.50% w/v    -   Carbopol® 940 at a concentration of about 1.5% w/v;    -   BAK at a concentration of about 0.015% w/v; and    -   phosphate buffer at a concentration of about 3 mM,    -   wherein the pH is about 5.25.

In another embodiment, the ophthalmological composition comprises:

-   -   aceclidine at a concentration of about 1.75% w/v;    -   mannitol at a concentration of about 2.5% w/v;    -   polysorbate 80 at a concentration of about 0.25% w/v;    -   NaCl at a concentration of about 0.1% w/v;    -   boric acid at a concentration of about 0.12% w/v;    -   Carbopol® 940 at a concentration of about 0.95% w/v; and    -   BAK at a concentration of about 0.015% w/v;    -   wherein the pH is about 5.

In another embodiment, the ophthalmological composition comprises:

-   -   aceclidine at a concentration of about 1.75% w/v;    -   mannitol at a concentration of about 2.5% w/v;    -   polysorbate 80 at a concentration of about 0.50% w/v;    -   NaCl at a concentration of about 0.05% w/v;    -   boric acid at a concentration of about 0.2% w/v;    -   Carbopol® 940 at a concentration of about 0.95% w/v;    -   BAK at a concentration of about 0.01% w/v; and    -   phosphate buffer at a concentration of about 3 mM,    -   wherein the pH is about 5.

In another embodiment, the ophthalmological composition comprises:

-   -   aceclidine at a concentration of about 1.75% w/v;    -   mannitol at a concentration of about 2.5% w/v;    -   polysorbate 80 at a concentration of about 0.1% w/v;    -   boric acid at a concentration of about 0.2% w/v;    -   Carbopol® 940 at a concentration of about 0.9% w/v;    -   BAK at a concentration of about 0.05% w/v; and    -   phosphate buffer at a concentration of about 3 mM,    -   wherein the pH is about 5.

In another embodiment, the ophthalmological composition comprises:

-   -   aceclidine at a concentration of about 1.75% w/v;    -   mannitol at a concentration of about 2.5% w/v;    -   polysorbate 80 at a concentration of about 0.1% w/v;    -   NaCl at a concentration of about 0.1% w/v;    -   boric acid at a concentration of about 0.12% w/v;    -   Carbopol® 940 at a concentration of about 0.95% w/v;    -   BAK at a concentration of about 0.01% w/v; and    -   phosphate buffer at a concentration of about 3 mM,    -   wherein the pH is about 5.

In another embodiment, the ophthalmological composition comprises:

-   -   aceclidine at a concentration of about 1.75% w/v;    -   tropicamide at a concentration of about 0.01% w/v;    -   mannitol at a concentration of about 2.5% w/v;    -   polysorbate 80 at a concentration of about 5.0% w/v;    -   CMC at a concentration of about 1.4% w/v;    -   BAK at a concentration of about 0.015% w/v; and    -   phosphate buffer at a concentration of about 3 mM,    -   wherein the pH is about 5.

In another embodiment, the ophthalmological composition comprises:

-   -   aceclidine at a concentration of about 1.75% w/v;    -   tropicamide at a concentration of about 0.02% w/v;    -   mannitol at a concentration of about 2.5% w/v;    -   polysorbate 80 at a concentration of about 0.25% w/v;    -   NaCl at a concentration of about 0.1% w/v;    -   boric acid at a concentration of about 0.12% w/v;    -   Carbopol® 940 at a concentration of about 0.95% w/v; and    -   BAK at a concentration of about 0.01% w/v.    -   wherein the pH is about 5.

In another embodiment, the ophthalmological composition comprises:

-   -   aceclidine at a concentration of about 1.75% w/v;    -   tropicamide at a concentration of about 0.015% w/v;    -   mannitol at a concentration of about 2.5% w/v;    -   polysorbate 80 at a concentration of about 0.75% w/v;    -   NaCl at a concentration of about 0.05% w/v;    -   boric acid at a concentration of about 0.2% w/v;    -   Carbopol® 940 at a concentration of about 0.95% w/v;    -   BAK at a concentration of about 0.01% w/v; and    -   phosphate buffer at a concentration of about 3 mM.    -   wherein the pH is about 5.

In another embodiment, the ophthalmological composition comprises:

-   -   aceclidine at a concentration of about 1.75% w/v;    -   tropicamide at a concentration of about 0.025% w/v;    -   mannitol at a concentration of about 2.5% w/v;    -   polysorbate 80 at a concentration of about 0.1% w/v;    -   boric acid at a concentration of about 0.2% w/v;    -   Carbopol® 940 at a concentration of about 0.9% w/v;    -   BAK at a concentration of about 0.05% w/v; and    -   phosphate buffer at a concentration of about 3 mM.    -   wherein the pH is about 5.

In another embodiment, the ophthalmological composition comprises:

-   -   aceclidine at a concentration of about 1.75% w/v;    -   tropicamide at a concentration of about 0.02% w/v;    -   mannitol at a concentration of about 2.5% w/v;    -   polysorbate 80 at a concentration of about 0.1% w/v;    -   NaCl at a concentration of about 0.1% w/v;    -   boric acid at a concentration of about 0.12% w/v;    -   Carbopol® 940 at a concentration of about 0.95% w/v;    -   BAK at a concentration of about 0.01% w/v; and    -   phosphate buffer at a concentration of about 3 mM.    -   wherein the pH is about 5.

In another embodiment, the ophthalmological composition comprises:

-   -   aceclidine at a concentration of about 1.75% w/v;    -   tropicamide at a concentration of about 0.040% w/v;    -   polyoxyl 40 stearate at a concentration of about 5.0% w/v;    -   mannitol at a concentration of about 2.5% w/v;    -   acetate or phosphate buffer at a concentration of about 3.0 mM;        and    -   BAK at a concentration of about 0.01% w/v,    -   wherein said composition has a pH of about 4.75.

In another embodiment, the ophthalmological composition comprises:

-   -   aceclidine at a concentration of about 1.55% w/v;    -   tropicamide at a concentration of about 0.040% w/v;    -   polyoxyl 40 stearate at a concentration of about 5.0% w/v;    -   citric acid monohydrate at a concentration of about 0.1% w/v;    -   mannitol at a concentration of about 4.0% w/v;    -   Carbopol® 940 at a concentration of 0.09% w/v; and    -   acetate or phosphate buffer at a concentration of about 3.0 mM;    -   wherein said composition has a pH of about 5.0.

In another embodiment, the ophthalmological composition comprises:

-   -   aceclidine at a concentration of about 1.50% w/v;    -   tropicamide at a concentration of about 0.042% w/v;    -   polyoxyl 40 stearate at a concentration of about 5.5% w/v;    -   mannitol at a concentration of about 2.5% w/v;    -   phosphate buffer at a concentration of about 3.0 mM;    -   Carbopol® 940 at a concentration of about 0.85% w/v; and    -   BAK at a concentration of about 0.01% w/v,    -   wherein said composition has a pH of about 4.75.

In another embodiment, the ophthalmological composition comprises:

-   -   aceclidine at a concentration of about 1.45% w/v;    -   tropicamide at a concentration of about 0.042% w/v;    -   polyoxyl 40 stearate at a concentration of about 5.5% w/v;    -   citric acid monohydrate at a concentration of about 0.1% w/v;    -   acetate buffer at a concentration of about 3.0 mM; and    -   Carbopol® 940 at a concentration of about 0.75% w/v,    -   wherein said composition has a pH of about 4.75.

In another embodiment, the ophthalmological composition comprises:

-   -   aceclidine at a concentration of about 1.45% w/v;    -   tropicamide at a concentration of about 0.042% w/v;    -   polyoxyl 40 stearate at a concentration of about 5.5% w/v;    -   mannitol at a concentration of about 2.0% w/v;    -   citric acid monohydrate at a concentration of about 0.1% w/v;    -   phosphate buffer at a concentration of about 3.0 mM; and    -   Carbopol® 940 at a concentration of about 1.0% w/v,    -   wherein said composition has a pH of about 4.75.

In another embodiment, the ophthalmological composition comprises:

-   -   about 1.75% w/v aceclidine;    -   about 2.5% w/v mannitol;    -   about 2.75% w/v polysorbate 80; and    -   about 1.25%; 1.0%-1.80% w/v hydroxypropylmethyl cellulose        (depending on its molecular weight).

In another embodiment, the ophthalmological composition comprises:

-   -   about 1.75% w/v aceclidine;    -   about 0.005% to about 0.011% tropicamide;    -   about 2.5% w/v mannitol;    -   about 2.75% w/v polysorbate 80; and    -   about 1.25%; 1.0%-1.80% w/v hydroxypropylmethyl cellulose        (depending on its molecular weight).

In another embodiment, the ophthalmological composition comprises:

-   -   about 1.75% w/v aceclidine;    -   about 0.010% w/v tropicamide;    -   about 2.5% w/v mannitol;    -   about 5.0% w/v polysorbate 80;    -   about 1.40% w/v carboxymethyl cellulose high viscosity;    -   about 3 mM phosphate buffer; and    -   about 0.010% BAK=as preservative,    -   with a pH of about 5.0.

In another embodiment, the ophthalmological composition comprises:

-   -   about 1.75% w/v aceclidine;    -   about 0.006% w/v tropicamide;    -   about 2.5% w/v mannitol;    -   about 2.5% w/v polysorbate 80;    -   about 1.25%; 1.0%-1.80% w/v hydroxypropylmethyl cellulose        (depending on its molecular weight);    -   about 3 mM phosphate buffer; and    -   about 0.020% BAK=as preservative,    -   with a pH of about 5.0.

In another embodiment, the ophthalmological composition comprises:

-   -   about 1.75% w/v aceclidine;    -   about 0.006% w/v tropicamide;    -   about 2.5% w/v mannitol;    -   about 2.5% w/v polysorbate 80;    -   about 1.25%; 1.0%-1.80% w/v hydroxypropylmethyl cellulose        (depending on its molecular weight);    -   about 3 mM phosphate buffer;    -   about 0.50% w/v NaCl; and    -   about 0.020% BAK=as preservative,    -   with a pH of about 5.0.

In another embodiment, the ophthalmological composition comprises:

-   -   about 1.75% w/v aceclidine;    -   about 2.5% w/v mannitol;    -   about 3.5% w/v polysorbate 80;    -   about 1.25%; 1.0%-1.80% w/v hydroxypropylmethyl cellulose        (depending on its molecular weight);    -   about 3 mM phosphate buffer;    -   about 0.50% w/v NaCl; and    -   about 0.020% BAK or 0.15% sorbic acid as preservative,    -   with a pH of about 5.0.

In another embodiment, the ophthalmological composition comprises:

-   -   about 1.75% w/v aceclidine;    -   about 2.5% w/v mannitol;    -   about 3.5% w/v polysorbate 80; and    -   about 1.25%; 1.0%-1.80% w/v hydroxypropylmethyl cellulose        (depending on its molecular weight);

In another embodiment, the ophthalmological composition comprises:

-   -   about 1.75% w/v aceclidine;    -   about 2.5% w/v mannitol;    -   about 3.5% w/v polysorbate 80;    -   about 1.25%; 1.0%-1.80% w/v hydroxypropylmethyl cellulose        (depending on its molecular weight); and    -   one or more excipient selected from the group consisting of        about 0.50% w/v sodium chloride, about 0.02% w/v benzalkonium        chloride, about 0.10% w/v sorbate, about 0.01% w/v        ethylenediaminetetraacetic acid (EDTA) and 0.10% w/v citric        acid.

In another embodiment, the ophthalmological composition comprises:

-   -   about 1.75% w/v aceclidine;    -   about 2.5% w/v mannitol;    -   about 0.01% w/v tropicamide;    -   about 0.1% w/v sodium citrate, anhydrous;    -   about 0.02% w/v benzalkonium chloride;    -   about 0.12% w/v sorbic acid;    -   about 0.1% w/v disodium edetate dihydrate;    -   about 4.0% w/v polysorbate 80; and    -   about 1.25% w/v hydroxypropylmethyl cellulose,    -   wherein the pH is about 5.0.

In another embodiment, the ophthalmological composition comprises:

-   -   about 1.75% w/v aceclidine;    -   about 2.5% w/v mannitol;    -   about 0.01% w/v tropicamide;    -   about 0.1% w/v sodium citrate, anhydrous;    -   about 0.02% w/v benzalkonium chloride;    -   about 0.1% w/v sorbic acid;    -   about 0.1% w/v EDTA;    -   about 3.5% w/v polysorbate 80; and    -   about 1.25%; 1.0%-2.25% w/v hydroxypropylmethyl cellulose        (depending on its molecular weight),    -   wherein the pH is about 5.0.

In another embodiment, the ophthalmological composition comprises:

-   -   about 1.75% w/v aceclidine;    -   about 2.5% w/v mannitol;    -   about 0.01% w/v tropicamide;    -   about 3 mM phosphate buffer;    -   about 0.02% w/v benzalkonium chloride;    -   about 0.1% w/v sorbic acid;    -   about 0.10% w/v citrate;    -   about 3.5% w/v polysorbate 80; and    -   about 1.25%; 0.25%-2.25% w/v hydroxypropylmethyl cellulose        (depending on its molecular weight);    -   wherein the pH is about 5.0.

In another embodiment, the ophthalmological composition comprises:

-   -   aceclidine at a concentration of 1.5% w/v, mannitol at a        concentration of 2.5% w/v.

In another embodiment, the ophthalmological composition comprises:

-   -   aceclidine at a concentration of 1.55% w/v, mannitol at a        concentration of 2.5% w/v.

In another embodiment, the ophthalmological composition comprises:

-   -   aceclidine at a concentration of 1.6% w/v, mannitol at a        concentration of 2.5% w/v.

In another embodiment, the ophthalmological composition comprises:

-   -   aceclidine at a concentration of 1.65% w/v, mannitol at a        concentration of 2.5% w/v.

In another embodiment, the ophthalmological composition comprises:

-   -   aceclidine at a concentration of 1.7% w/v, mannitol at a        concentration of 2.5% w/v.

In another embodiment, the ophthalmological composition comprises:

-   -   aceclidine at a concentration of 1.75% w/v, mannitol at a        concentration of 2.5% w/v.

In another embodiment, the ophthalmological composition comprises:

-   -   aceclidine at a concentration of 1.80% w/v, mannitol at a        concentration of 2.75% w/v and Carbopol® 940 at a concentration        of 0.09% w/v.

In another embodiment, the ophthalmological composition comprises:

-   -   aceclidine at a concentration of 1.48% w/v, mannitol at a        concentration of 1.5% w/v and Carbopol® 940 at a concentration        of 0.50% w/v.

In another embodiment, the ophthalmological composition comprises:

-   -   aceclidine at a concentration of 1.80% w/v, mannitol at a        concentration of 2.5% w/v and Carbopol® 940 at a concentration        of 0.9% w/v.

In certain preferred embodiments, the present invention is directed tocompositions for the treatment of presbyopia comprising about 1.75% w/vaceclidine, about 4.0% w/v polysorbate 80, about 2.5% w/v mannitol,about 1.2% w/v hydroxypropylmethyl cellulose, about 0.1% w/vethylenediaminetetraacetic acid, about 0.02% w/v benzalkonium chloride,about 0.12% w/v potassium sorbate and about 0.077% w/v citrate, whereinthe composition has a pH of about 5.0.

In certain preferred embodiments, the present invention is directed tocompositions for the treatment of presbyopia comprising about 1.75% w/vaceclidine, about 4.0% w/v polysorbate 80, about 2.5% w/v mannitol,about 1.2% w/v hydroxypropylmethyl cellulose, about 0.1% w/vethylenediaminetetraacetic acid, about 0.02% w/v benzalkonium chloride,about 0.12% w/v potassium sorbate and about 0.1% w/v citrate, whereinthe composition has a pH of about 5.0.

In certain preferred embodiments, the present invention is directed tocompositions for the treatment of presbyopia comprising about 1.40% w/vaceclidine, about 2.0% w/v polyoxyl stearate, about 2.50 w/v mannitol,about 0.10% w/v ethylenediaminetetraacetic acid, about 0.02 w/vbenzalkonium chloride, about 0.12 w/v potassium sorbate and about 0.1w/v citrate, wherein the composition has a pH of about 5.0.

In another embodiment the ophthalmological compositions are those inTables 1 and 2.

TABLE 1 Representative Aceclidine Compositions Formula A B C D E F G H IAceclidine 1.75% 1.35% 1.40% 2.40% 1.50% 1.00% 0.75% 0.55% 1.50%Polysorbate 80 — 1.00% — 2.00% 1.00% 0.25% 4.00% 4.00% — Poloxamer 407 —— — — 0.10% 1.00% — — — Poloxamer 188 — — — 2.00% 0.10% 0.25% — — 1.00%Cyclodextrin — — 1.00% — 0.50% 0.25% — 1.00% 1.00% CMC — — 0.75% 0.85%1.00% 0.80% 1.10% 1.20% 0.25% (2% = 3500 cps) NaCl 0.90% 0.80% 0.90%0.75% 0.75% 0.85% 0.90% 0.90% 0.90% K Sorbate 0.10% 0.10% 0.10% 0.12%0.12% 0.10% 0.10% 0.10% 0.10%

TABLE 2 Representative Aceclidine Compositions Formula J K L Aceclidine1.75% 1.75% 1.75% Tropicamide 0.010%  0.010%  0.006%  Mannitol — 2.00%2.50% Polysorbate 80 1.00% 2.00% 4.00% Poloxamer 407 0.50% 0.20% —Poloxamer 188 — 0.20% — Cyclodextrin — 0.20% — CMC 1.00% 0.85% 1.30% (2%= 3500 cps) NaCl 0.90% 0.65% 0.90% K Sorbate 0.10% 0.10% 0.10%

The following Examples are provided solely for illustrative purposes andare not meant to limit the invention in any way.

EXAMPLES Example 1 Effect of Aceclidine on Vision of Subjects Aged 47 to67 Years

Table 3 demonstrates the effect on the near focus ability of presbyopicsubjects before and after ophthalmological administration of acomposition containing aceclidine. Each composition included aceclidinein the concentrations indicated and 5.5% w/v HPβCD, 0.75% w/v CMC, 0.25%w/v NaCl and 0.01% w/v BAK. Additionally, compositions administered tosubjects 4 and 5 included 0.125% w/v tropicamide. As aceclidine is anenantiomer, the clinical effectiveness may vary with different ratios.For the present studies a nearly exact 50:50 ratio of stereoisomers wasmeasured as best determined by polarimetry.

TABLE 3 Effects of aceclidine on vision of presbyopic patients.Aceclidine Vision Baseline Post Gtt 15″ Date # Age % R Pre Dist L PreDist R Pre Near L Pre Near R Post Dist L Post Dist R Post Near L PostNear Effect (h) 8/21/2013: 1 67 1.5 20.20 20.30 20.60 20.60 20.20 20.2020.15 20.15 9.00 8/22/2013 2 52 1.5 20.30 20.30 20.50 20.50 20.25 20.2520.25 20.20 8.00 8/23/2013 3 61 1.5 20.40 20.30 20.60 20.50 20.20 20.2520.15 20.15 8.00 8/23/2013: 4 61 1.1 20.20 20.25 20.80 20.50 20.15 20.1520.20 20.15 12.00 8/23/2013: 5 53 1.1 20.20 20.20 20.60 20.60 20.2020.20 20.25 20.25 7.00 8/24/2013: 6 47 1.5 20.25 20.25 20.100 20.10020.20 20.20 20.15 20.15 8.00 8/25/2013 7 58 1.5 20.30 20.200 20.10020.30 20.25 20.30 20.20 20.30 8.00

As seen in Table 3 all subjects had less than perfect near vision(20.20) in both the left and right eye (object at 15 inches from theeye) and most subjects had less than perfect distance vision beforeadministration of the composition. After administration of thecomposition all subjects experienced an improvement in their near visionthat lasted from 7 to 12 hours. Surprisingly, the majority of subjectsalso experienced improvement of their distance vision for the same timeperiod. Still more surprisingly the improvement in near point was muchcloser than 16″ typically required for comfortable reading, in somecases to about 8.5″ more commonly seen in individuals 30 or less. Theaddition of tropicamide, a cycloplegic agent, had no additive ordeleterious effect on vision correction.

Example 2 Effect of Concentration of Concentration of Aceclidine andTropicamide

TABLE 4 Effect of concentration of concentration of aceclidine andtropicamide. #1 #2 #3 #4 #5 (OD) #5 (OS) #6 #7 Brimonidine 0.03% 0.03%0.03% 0.03% 0.03% 0.03% 0.03% Poloxamer 407 5.5% HPBCD 5.5% 5.5% 5.5%55% 55% 5.5% 5.5% Aceclidine 1.5% 1.5% 0.75% 1.1% 11% 11% 1.1% 1.1%Tropicamide 0.014% 0.021% 0.028% 0.042% 0.062% NaCl 0.25% 0.25% 0.25%0.25% 0.25% 0.25% 0.25% 0.25% CMC 0.75% 0.75% 0.75% 0.75% 0.25% 0.75%0.75% 0.75% BAK 0.1% 0.1% 0.1% 0.1% 0.1% 0.1% 0.1% 0.1% Redness (15 m)3+ 1 0.5 0.5 0 0 0 0 Redness (30 m) 1.5 0.5 0.25 0.25 0 0 0 0 Brow Ache(60 m) 2+ 2+ 2 0.5 0.5 0.0 0.0 0.0 Stinging (10 m) 2 2 0.5 0 0 0 0 0BD-OD 20.20 20.20 20.20 20.20 20.20 20.20 20.20 20.20 BD-OS 20.25 20.2520.25 20.25 20.25 20.25 20.25 20.25 BN-OD 8 pt 8 pt 8 pt 8 pt 8 pt 8 pt8 pt 8 pt BN-OS 7 pt 7 pt 7 pt 7 pt 7 pt 7 pt 7 pt 7 pt BP photopic 3 mm3 mm 3 mm 3 am 3 mm 3 mm 3 mm 3 mm BP-mesopic 5 mm 5 mm 5 mm 5 mm 5 mm 5mm 5 mm 5 mm Miosis start (m) 15 15 15 15 15 15 15 Miosas (OU) (1 hr)1.63 mm 1.63 mm 2025 mm 1.03 mm 1.63 cm 1.63 mm 1.63 mm 1.70 mm Distance(OU) (20 m) 20.20 20.20 20.20 20.20 20.20 20.20 20.20 20.20 Distance(OD) (1 hr) 20.15 + 2 20.15 + 2 20.20 20.15 + 2 20.15 + 2 20.15 + 220.15 + 2 20.15 + 2 Distance (OS) (1 hr) 20.15 + 2 20.15 + 2 20.2020.15 + 2 20.15 + 2 20.15 + 2 20.15 + 2 20.15 + 2 Distance (OU) (1 hr)20.10-3 20.10.3 20.15 20.10-3 20.10.3 20.10-3 20.10.3 20.10.3 Near (OU)(20 m) 4 pt 4 pt 4pt 4 pt 4pt 4 pt 4 pt 4 pt Time (hr) 12.5 12.5 6.5 1110 10

Abbreviations: (C) indicates corrected vision, (in) indicates minutes,(hr) indicates hour, mm indicates millimeters, BD indicates baselinedistance vision; BN indicates baseline near vision, BP indicatesbaseline pupil size, GD indicates right eye; OS indicates left eye andOU indicates both eyes.

All percentages are w/v. “pt” reflects size of print materials, 4 beingequivalent to 20/20 vision and 3 to 20/15 vision.

“Time” refers to duration of the effect.

As seen in Table 4 aceclidine at a concentration of at least 1.1% w/vwas able to reduce the size of the pupil to 1.63 mm 1 hour after topicalinstillation resulting in corrected near and distance vision for atleast 10 hours. Lowering of the concentration of aceclidine to 0.75% w/v(formula #3) reduced the miotic effect to 2.0-2.5 mm after 1 hour andvision correction lasted only 6.5 hours. The addition of 0.03% w/vbrimonidine reduced redness of the eye (4 out of 4 without brimonidine,not shown) to 1.5 out of 4 within 30 minutes after topical instillationwhich was maintained for the entire time vision was corrected. Switchingthe nonionic surfactant to HPβCD (formulas #2-6) further reduced theredness of the eye. Lowering of the concentration of aceclidine to 0.75%w/v (formula #3) further reduced eye redness but as mention above alsoreduced the vision correction duration of the formula.

A brow ache and stinging in the eye were noticeable in formulas #1-3with a 2 out of 4 level of pain which was also associated with feelingsof slight nausea, upset stomach and fatigue. Surprisingly, the additionof a cycloplegic agent, tropicamide, reduced brow ache and stinging to0.5 out of 4 and 0 out of 4 respectively with brow ache dissipatingafter 60 minutes (formula #4). Further, the raising of the concentrationof aceclidine to 1.1% w/v restored the longer duration of correctedvision seen in formulas #1-2 without increasing eye redness. However,upon re-topical instillation of formula #4 at the end of the 10 hoursnoticeable brow ache occurred. Topical instillation of formula #5 (OD)and (OS), with increased tropicamide concentrations, following formula#4 relieved the brow ache experienced with re-installation of formula#4. Upon a₃rd topical instillation, at the end of the effective durationof formula #5, re-topical instillation of formula #5 again led toconsiderable brow ache. Once again, in formula #6, raising theconcentration of tropicamide was able to overcome the brow ache.Additionally, and unexpectedly, tropicamide, despite being a cycloplegicagent, had no effect on pupil miosis or vision correction. Surprisingly,the addition of tropicamide resulted in a prolonged duration of optimalpupil size constriction.

To determine the effect of brimonidine on pupil miosis, formula #7, wasadministered. Administration of formula #7 resulted in only a slightdecrease in pupil miosis to 1.70 mm with identical distance and nearvision improvement to that of formula #5. A 2-3+ conjunctival injectionwas noted.

All baseline vision data was based on vision corrected with distancecontact lenses. Near vision was noted by subject as outstanding from 8inches to the horizon at 1.5 hours after installation. A MarcoAutorefractor with infrared camera and superimposed pupil calibrationscale was used for all pupil size measurements. Once an image wasselected it remained on screen allowing accurate calibration.

Example 3 Effect of Concentration of Aceclidine, Brimonidine,Guanfacine, Fadolmidine, Tropicamide and Additives

TABLE 5 Effect of concentration of aceclidine, brimonidine, guanfacine,fadolmidine, tropicamide and additives. AB2T AB4T AB6T AB11T AB12TPROPH13 Aceclidine 1.55 1.55 1.55 1.55 1.85 1.55 Brimonidine 0.037 0.0370.037 0.037 Fadoknidine 0.037 Guanfacine 0.037 HPBCD 5.5 S.S 5.5 5.5 5.55 Tropicamide 0.043 0.043 0.043 0.043 0.042 0.043 CMC* 0.075 0.075 0.0750.075 0.075 0.075 NaCl 0.025 0.025 0.02$ 0.025 0.025 0.025 BAK 0.01 0.010.01 0.01 0.01 0.01 Glycerin 0.1 0.1 0.1 Poloxamer 188 0.1 0.05 Polyoxyl40 stearate 0.05 pH 6.5 7.5 7.5 7.5 7.0 7.5 nasal congestion 0 0 0 0 0 0stinging initial 0.75 0 1.5 3.5 0 1.5 stinging 3 mm 0.5 0 0 wash out 0 0redness initial 0 0 1 D/C 1 1 redness 15 min 0 0 0 D/C 0 0 whitening 0 00 D/C 1.5 1.5 pain 0 0 0 D/C 0 0 vision near 20.30 20.15 20.15 D/C 20.1520.15 vision distance 20.20 20.20 20.20 D/C 20.20 20.20 onset (min) 2012 16 D/C 12 16 duration (hrs) 5.5 7.5 7.5 D/C 7.5 7.5 color clearyellow yellow yellow yellow yellow OVERALL 2.5 3.9 3.8 0 4 3.9

*1%=2,500 cps

All percentages are w/v. Scores for nasal congestion, stinging initial,stinging, 3 min, redness initial, redness 15 min, whitening, pain andoverall are out of 4.

“pt” reflects size of print materials, 4 being equivalent to 20/20vision and 3 to 20/15 vision.

Baseline vision was 20.20 both eyes for distance; 20.70 right eyeunaided for near; 20.80 left eye for near (best @ 16″).

D/C stands for discontinued after eye washing due to intolerablestinging.

Aceclidine at a concentration of 1.55% w/v was able to reduce the sizeof the pupil to about 1.63 mm 30 minutes after topical instillationresulting in corrected near and distance vision to 20.20 or better forat least 6 hours, with noticeable affect lasting about 7.5 hours as seenin Table 5. Lowering of the concentration of aceclidine to 1.25% w/v(not shown) resulted in useful near vision improvement to about20.25-20.30, but not as effective as at the higher dose range alkalinepH resulted in quicker onset, longer duration, and greater effect. Theaddition of 0.037% w/v brimonidine reduced redness of the eye (4 out of4 without brimonidine, not shown) to baseline within 15 minutes aftertopical instillation which was maintained for the about the entire timevision was corrected. Adding glycerin 0.10% w/v noticeably reducedstinging. Adding instead poloxamer 188 0.05% w/v and polyoxyl 40stearate 0.05% w/v however reduced initial stinging further but was moreviscous. The combination of glycerin 0.1% w/v, poloxamer 188 0.1% w/v ata pH of 6.5 was noticeably reduced in onset, duration, comfort andeffectiveness. AB11T did not include glycerin, poloxamer 188, orpolyoxyl 40 stearate, which resulted in substantial stinging anddiscontinuation of the experiment with eye flush irrigation immediatelyafter topical instillation. Substitution of guanfacine 0.037% w/v inAB12T for brimonidine resulted in minimal initial redness with prolongedredness reduction and some degree of whitening, and appeared to provideoverall the best cosmesis though requiring slightly higher aceclidineconcentration for optimal effect.

All baseline vision data was based on vision corrected with distancecontact lenses. Near vision was noted by subject as outstanding from 8to 10 inches to the horizon at 30 minutes after installation for AB4Tand AB6T.

AB4T and AB6T were repeated both monocularly and binocularly.Substantial improvement in depth perception, near point acuity to 3 pt(20.15), and near point distance (8″, 20.20) was noted when both eyeswere treated vs. monocular treatment. Monocular treatment resulted inworsening of vision with both eyes open versus testing only the treatedeye.

Example 4 Effect of Concentration of Aceclidine, Brimonidine,Tropicamide, and Additives

TABLE 6 Effect of concentration of aceclidine, brimonidine, tropicamide,and additives. #8 #9 #10 #11 #12 #13 #14 Aceclidine 1.61% 1.61% 1.61%1.61% 1.61% 1.53% 1.53% Tropicamide 0.042% 0.042% 0.042% 0.042% 0.042%0.044% 0.044% Brimonidine 0.042% 0.042% 0.042% 0.042% 0.042% CMC 0.75%0.75% 0.80% 0.87% 0.75% 0.75% 0.75% NaCI 0.25% 0.25% 0.50% 0.50% 0.25%0.50% 0.50% BAK 0.01% 0.01% 0.01% 0.01% 0.01% 0.01% 0.01% PH 7.00 7.007.00 7.00 8.00 7.00 7.00 phosphate buffer 5 mM 5 mM 5 mM 6 mM 5 mM 5 mMborate buffer 5 mM Onset (min) 15 15 15 15 15 15 15 Duration (hrs) 710-12 10-12 10-12 7 9 7 Pupil range (mm) 1.5-1.7 1.5-1.7 1.5-1.7 1.5-1.71.5-1.7 1.8-2.0 1.8-2.0 Dimming 0-4 1.5 1.5 1.5 1.5 1.5 0.5 0.5 Sting0-4 1 1 1 1 1 1 1 Ache 0-4 0.25 0.25 0.25 0.25 0.25 0.00 0.00 Redness0-4 0.5 0.5 0.5 0.5 1.5 1.0 0.5 Other watery watery sl thicker slresidue watery watery watery Overall 0-5 3.5 3.5 4 4 2.5 4.5 4.75 #15#16 #17 #18 #19 #20 #21 Aceclidine 1.53% 1.53% 1.53% 1.53% 1.45% 1.65%1.75% Tropicamide 0.044% 0.044% 0.044% 0.044% 0.042% 0.044% 0.035%Brimonidine 0.042% 0.042% 0.042% 0.042% 0.042% 0.042% CMC 0.80% 0.80%0.80% 0.80% 0.75% 0.75% 0.75% NaCI 0.50% 0.75% 0.75% 1.00% 0.25% 0.25%0.25% BAK 0.01% 0.01% 0.01% 0.01% 0.01% 0.01% 0.01% PH 7.00 7.00 8.007.00 7.00 7.00 8.00 phosphate buffer 5 mM 5 mM 5 mM 5 mM 5 mM 5 mM 6 mMborate buffer Onset (min) 15 15 15 15 15 15 15 Duration (hrs) 10-12 9 97 7 7 7 Pupil range (mm) 1.8-2.0 1.8-2.0 1.8-2.0 1.8-2.0 1.8-2.1 1.8-2.11.8-2.2 Dimming 0-4 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Sting 0-4 1 1 1 1 1 1 1Ache 0-4 0.00 0.00 0.00 0.00 0.00 0.25 0.00 Redness 0-4 0.5 0.5 1.0 0.50.5 0.5 0.5 Other sl thicker sl thicker sl thicker thicker watery waterywatery Overall 0-5 5 5 5 4 4 4 4

As seen in Table 6 formulas #8-9 an increase in brimonidine to 0.42% w/vresulted in redness reduction to 0.5 while 0.750 w/v CMC resulted in awatery consistency Unexpectedly, increasing CMC from 0.75% w/v to arange of 0.800% w/v to 0.87% w/v and increasing NaCl from 0.25% w/v to0.75% w/v in formulas #10-11 resulted in a thicker consistency and anincreased residence time from 7 hours to 10-12 hours and decreased theamount of drug that drained into the nasolacrimal duct. This decreaseddrug delivery to the nasal passages results in less nasal congestion.

In formulas #13-18 a decrease in the amount of aceclidine from 1.61% to1.53% w/v resulted in a pupil size range from 1.8-2.0 mm. Dimming as aresult of the restriction of the pupil decreased linearly from 1.5 to0.5 with the decreased amount of aceclidine. Specifically, the 1.8 to2.0 mm pupil created 41% more light than the 1.5 to 1.7 mm pupil.Surprisingly, the 1.8 to 2.0 mm pupil had a near depth increase of 1.75D. This is only a 0.25 D loss from the beneficial 2.00 D seen with the1.5-1.7 mm range. Thus, the 1.80 to 2.0 mm range produces 4100 morelight while still allowing the full benefit of increased near vision inindividuals under 60 years of age; whereas, individuals 60 years of ageand over still experience total computer benefit and some increased nearbenefit.

The increase in tropicamide concentration from 0.042% w/v (formulas#8-#11) to 0.044% w/v (formulas #13-#18) resulted in a decrease in acheto negligible amounts. The amount of ache may also be correlated withthe age of the individual. For those individuals under the age of 45, anincrease of tropicamide concentration to a range from 0.046% to 0.060%w/v may be preferred.

Further, Table 6 shows an unexpected result seen in formulas #13 and #17where the increase of NaCl from 0.25% w/v to a range of 0.50 to 0.75%w/v resulted in an acceptable redness score of only 1.0 even without theaddition of the redness reducing agent brimonidine.

Formulas #15, #16 and #17 each result in an overall maximum rating of 5by combining the benefits of: (1) reduced aceclidine concentrations toimprove the amount of light produced without significantly affecting thenear vision benefits seen in formulas #8-#12; (2) increased NaClconcentrations resulting in a further reduction in redness even in theabsence of brimonidine; and (3) increased CMC concentrations resultingin longer residency time on the eye.

Formula #19 is an excellent alternative for the minority of individualsthat are high responders to formulas #15-#17 and get noticeable dimmingwith 1.53% w/v aceclidine. Formula #20 is an excellent alternative forthe minority of individuals that are low responders to formula #19.Lastly, Formula #21 is an excellent alternative for the minority ofindividuals that are low responders and get poor pupil response withFormula #20.

Example 5 Comparison of Effects of Polyoxyl 40 Stearate, HPβCD andPoloxamer 407

TABLE 7 Comparison of Effects of Polyoxyl 40 Stearate, HPβCD andPoloxamer 407. #22 #23 #24 Aceclidine  1.45%  1.45%  1.45% Tropicamide0.044% 0.044% 0.044% Brimonidine 0.040% 0.040% 0.040% Polyoxyl 40Stearate  5.5% HPβCD  5.5% Poloxamer 407  5.5% CMC  0.80%  0.80%  0.80%NaCl 0.037% 0.037% 0.037% EDTA 0.015% 0.015% 0.015% BAK 0.007% 0.007%0.007% pH 7.00 7.00 7.00 phosphate buffer 5 mM 5 mM 5 mM NasalCongestion 0.00 0.50 1.50 Stinging 0.25 0.25 0.25 Wetting 4.00 4.00 4.00Redness 0.25 0.50 0.50 Visual Blur (<15 sec) 0.50 0.50 1.50 Duration 6-8hrs 6-8 hrs 6-8 hrs Overall 0-4 4.00 4.00 4.00

Clinical Protocol

20 presbyopic patients with full distance correction were each given oneof the above formulas (#22-#23). All patients received pre- andpost-drop distance and near acuity measurement, Zeiss Visante® (Visanteis a registered trademark of Carl Zeiss Meditec AG) optical adherencetomography, axial length and contrast acuity testing (i.e.Colenbrander-Michelson 10% Lum target) with the following results.

-   -   all patient achieved a miotic pupil of 1.5 to 2.20 mm;    -   no patient experienced ciliary ache, ciliary spasm, or induced        accommodation;    -   all patients achieved 20/30+ visual acuity or better at 14″ and        were very satisfied with their high contrast near vision results        and there was no significant complaint of burning or aching;    -   the duration of effect lasted 6-8 hrs in all cases;    -   binocular vision afforded all patients 1-1.5 additional lines of        near acuity over monocular testing; the last 10 patients were        tested at 20″ (i.e. computer distance, cell phone distance) and        all achieved 20/25 or better near visual acuity;    -   moderately hyperopic (approx. +2.25 sphere) uncorrected        presbyopes were very satisfied with distance visual acuity that        improved to a 20/25 or better level at distance and near vision        in the 20/30 range; and    -   uncorrected distance acuity was often improved for those        patients who chose not to routinely correct a small refractive        error.

As seen in Table 7, the use of polyoxyl 40 stearate provides the mostcomfortable aceclidine formulation with the least amount of visual blurand redness. To achieve similar results to that of formula #22, formula#23 requires 10-15% higher concentrations of the non-ionic surfactantand formula #24 requires 15-20% higher concentrations of the non-ionicsurfactant. HPβCD induced a color change over time, possibly indicativeof oxidation. Captisol® (sulfobutylether β-cyclodextrin) was substitutedwith similar findings.

Example 6 Modulation of Aceclidine Concentrations in a PreferredEmbodiment

Preferred embodiment:

-   -   Aceclidine 1.35%-1.55% w/v;    -   Polyoxyl 40 stearate 5.5% w/v;    -   NaCl 0.037% w/v;    -   a viscosity agent, preferably CMC 0.80% w/v or an amount of        Carbopol® 934 or 940 sufficient to achieve a viscosity of from        about 5 to about 35 cps upon topical instillation, such as        Carbopol® 940 at a concentration from about 0.09% to about 1.0%        w/v;    -   BAK 0.015% w/v; and    -   a phosphate, citrate, cerophosphate, or acetate buffer from        about 3 to about 10 mM,    -   wherein the pH is from about 4.75 to about 6.0.

For 1.35% w/v aceclidine—

-   -   Stinging on topical instillation 0.25/4.0 (lasting about 2-5        seconds);    -   Induced redness at 10 minutes: 1.0 to 1.5/4.0;    -   Induced redness at 30 minutes: 0.0 to 0.25/4.0;    -   Comfort: very high.    -   Wetting: very high, the eye maintaining sensation of improved        wetting for most of a 24-hour period after a single topical        instillation.    -   Depth of Focus distance: excellent.    -   Depth of Focus near: excellent.

In testing the above formulations on several subjects, it was discoveredthat there is a slight range in clinical effect depending on theconcentration of aceclidine, where 1.35%-1.55% w/v aceclidine ispreferred, but for which 1.35% w/v and 1.45% w/v confer the desiredbenefits on most subjects.

Further, it is discovered that the clinical effect of 1.35% w/vaceclidine can be improved when instilled as follows:

-   -   1) baseline effect: 1 drop to each eye.    -   2) enhanced effect: 2 drops to each eye.    -   3) greater effect: after 2) above repeat 1) above.

4) maximum effect: after 2) above repeat 2) above.

Example 7 Use of a Preferred Embodiment to Prolong Contact Lens Wear

Preferred embodiment:

Aceclidine 1.45% w/v;

-   -   Polyoxyl 40 stearate 5.5% w/v;    -   NaCl 0.037% w/v;    -   a viscosity agent, preferably CMC 0.80% w/v or an amount of        Carbopol® 934 or 940 sufficient to achieve a viscosity of from        about 5 to about 35 cps upon topical instillation, such as        Carbopol® 940 at a concentration from about 0.09% to about 1.0%        w/v; BAK 0.02% w/v; and    -   a phosphate, citrate, citrophosphate, or acetate buffer from        about 3 to about 10 mM,    -   wherein the pH is from about 4.75 to about 6.0.

As a baseline, the subject, who normally wore extended wear lenses (AirOptix®; Air Optix is a registered trademark of Novartis AG) for dailywear only, slept in these lenses overnight. On arising each morning, thesubject's vision was blurred, and the contact lenses required removaland cleaning of film and deposits that had formed overnight. Averagevision on arising at distance: 20.60; average vision at near on aMichelson contrast acuity chart: 20.80.

Then, for seven consecutive days the above formulation was instilledbetween 7 am and 10 am each day as a single dose. Subject wore the AirOptix® lenses throughout each day and slept in the lenses overnight.Upon arising each morning, the subject's vision at distance: 20.20+;vision at near 20.40 unaided (consistent with subject's baselinepresbyopia when the subject did not wear the lenses overnight andinstead inserted the lenses upon arising).

Example 8 Comparison of Effects of Polyoxyl 40 Stearate and Captisol®(Sulfobutylether β-Cyclodextrin

TABLE 8 Comparison of Effects of Polyoxyl 40 Stearate and Captisol®(sulfobutylether β-cyclodextrin). #25 #26 #27 #28 #29 *30 #31 #32 #33Aceclidine 1.35% 1.35% 1.35% 1.35% 1.35% 1.35% 1.35% 1.35% 1.35%Tropicamide 0.044% 0.044% 0.044% 0.044% 0.044% 0.044% 0.044% 0.044%0.044% Polvoxyl 40 stearate 5.5% 5.5% 5.5% 5.5% 5.5% 5.5% 5.5% Captisol®5.5% $.5% Cocamidopropyl betaine 0.10% EDTA 0.015% 0.015% 0.005% 0.005%0.005% 0.005% 0.015% CMC 1% = 2,500 cps 0.80% 0.80% 0.80% 0.80% 0.80%0.80% 0.80% 0.80% 0.80% NaCl 0.037% 0.037% 0.037% 0.037% 0.037% 0.037%0.037% 0.037% 0.037% Mannitol 4% 4% BAK 0.007% 0.007% 0.007% 0.007%0.007% 0.007% 0.007% 0.007% 0.007% Borate buffer (max) 4 4 4 4 4 4 4Phosphate buffer (md) 4 4 pH 7 7 7 7 7 7 7 7 7 Redness 10 min 1.25 1.252 2 1.75 1.75 0 0 0 Redness, 30 min 0 0 1.5 1.5 1.25 1.25 0 0 0 Pupil,30 min (mm) <2 <2 <2 <2 <2 <2 <2 <2 <3 Blur on instill (sec) 10 10 10 1010 10 10 10 10 Ache 0 0 0 0 0 0 1 0 0 Rating 4.00 4.00 2.00 2.00 2.502.50 1.00 5.00 TBD

As seen in Table 8, when using polyoxyl 40 stearate as the surfactantthe exclusion of EDTA results in reduced redness and best overall ratingamong polyoxyl 40 stearate compositions (Formulas #25 and #26). Theaddition of cocamidopropyl betaine (“CAPB”) further reduces rednesshowever results in significant ache (Formula #31). Replacing polyoxylstearate with Captisol® (sulfobutylether β-cyclodextrin) and addingmannitol achieves similar results in redness reduction as the additionof CAPB to polyoxyl 40 stearate but without the attendant ache resultingin the highest overall rating among aceclidine compositions (Formula#32). After several weeks formulations with Captisol® (sulfobutyletherβ-cyclodextrin) had an orange hue, possibly indicative of oxidation.

Example 9 Preferred Cold Chain Composition Composition

-   -   aceclidine at a concentration of about 1.40%-1.80% w/v; and    -   tropicamide at about 0.42% w/v;    -   polyoxyl 40 stearate at about 5.5% w/v;    -   mannitol at a concentration of about 2.5% to 4.5% w/v;    -   carbomer 940 at a concentration of about 0.09% to about 2.0%        w/v;    -   optionally, a preservative such as BAK at a concentration of        about 0.2% w/v;    -   optionally citrate at a concentration of about 0.1%;    -   optionally with acetate or phosphate buffer at 2-100 mM, more        preferably 3-5 mM    -   wherein said composition has a pH of about 4.50 to about 5.0;        and preferably, about 4.75 to about 5.0; and    -   wherein w/v denotes weight by volume

A composition as described above was administered to a 62-year-oldsubject. It resulted in pupils of 1.8-1.9 mm ou, 20.20+ reading vision,and 20.20+ distance vision; whereas without carbomer 940 reducedeffectiveness resulted at 2.5% mannitol, and no near vision effectresulted at 4.0% mannitol. No ciliary spasm or loss of distance visionresulted. Onset was within about 15 minutes. Transient redness of about1+/out of 4 was noted for about 20 minutes without alpha agonistvasoconstrictor. The presence or absence of BAK had no clinical effect,and was used to provide an optional preservative.

Example 10 Stabile Aceclidine Formulations Composition Tested:

-   -   aceclidine at a concentration of about 1.50% w/v;    -   tropicamide at a concentration of about 0.042% w/v;    -   polyoxyl 40 stearate at a concentration of about 5.5% w/v;    -   mannitol at a concentration of about 2.5% w/v;    -   citrate at a concentration of about 3 mM;    -   wherein said composition has a pH of about 4.75.

20 samples of the above composition were divided evenly and stored at25° C. and 4° C. Prior to storage, initial concentrations of aceclidinewere measured using high-pass liquid chromatography (“HPLC”). The amountof aceclidine in each solution was calculated by the area under theprincipal peak compared to a reference solution of aceclidine. Sampleswere then subject to storage for 3 months. Aceclidine measurements weretaken at 1, 2 and 3 months. Results of the stability test are shown inTable 9.

TABLE 9 Stability of Aceclidine in Cold Chain Storage 25° C. 4° C.Initial 100%  100%  1 month 92% 93% 2 months 75% 92% 3 months 50% 88%

As seen in Table 9 “cold chain storage” or storage of the aceclidinecomposition at from 2° C. to 8° C. resulted in a significant increase instability of aceclidine at all 3 time points.

Example 11 Use of Compositions Containing Little or No Cycloplegic Agent

Aceclidine alone causes incidence migraine-like severe ciliary spasm(brow ache) and myopic blur. These effects are inversely correlated toage with subjects age 40 reporting the highest incidence and subject age60+ reporting the lowest incidence. The addition of a cycloplegic agentreduces ciliary spasms and attendant brow ache, migranious headache,squeezing pressure around eyes or other symptoms of ciliary spasms. Theaddition of the cycloplegic agent, surprisingly, does not reduce themyopic effect of aceclidine. The addition of 2.5% w/v mannitol howeverdoes reduce the myopic effect of aceclidine. Increasing the aceclidineconcentration overcomes this reduction in myopic effect seen with theaddition of mannitol. Surprisingly, however, the increase in aceclidineis not coincident with an increase in ciliary spasm. Even moresurprising, the concentration of the cycloplegic agent can be reduced oreven eliminated in the presence of mannitol without an increase inciliary spasm. Thus, combining a higher concentration of aceclidine withlittle to no cycloplegic agent in the presence of mannitol results in animprovement of near vision acuity without attendant side effects on parwith lower concentrations of aceclidine and higher concentrations of thecycloplegic agent in the absence of a cycloplegic agent.

Further and unexpectedly, the addition of a nonionic surfactantincreases both the quantitative measure of near vision improvement andthe duration. This effect is concentration sensitive. In a preferredembodiment the non-ionic surfactant is at least 1%, preferably at least2%, more preferably from about 1% to about 5%, and most preferably about5%. For example, polysorbate 80 or polyoxyl 40 stearate at aconcentration from about 1% to about 5% w/v results in about 1.5 toabout 2.0 lines of improvement and a duration from about 4 to about 5hours.

Not to be held to particular theory, the increase in concentration of asurfactant may crowd the surface of the cornea, and at an optimalconcentration this crowding result in small and probably nanometerdiameters, which given the dual polarity of surfactants, where nonionicare most preferred, enhances corneal absorption of the entrapped highlypolar aceclidine molecules.

The further addition of a viscosity agent by itself does not enhanceduration. Surprisingly, the addition of a viscosity agent in aformulation with optimal ratios of aceclidine, tropicamide and anon-ionic surfactant dramatically improves duration. For example, aformulation of the present invention comprising 1.75% aceclidine, 2.5%mannitol, 0.01% tropicamide, 5% polysorbate 80 improves near vision in apresbyopic patient by up to 3 lines of vision acuity for about 4 toabout 5 hours. The addition of 1.4% CMC further increases the nearvision improvement to from about 7 to about 10 hours. Not to be held toa particular theory, a threshold above the critical micellar thresholdgreatly enhances permeation through the cornea by reducing micelle sizefrom micrometers to nanometers. See FIG. 2 .

Examples of compositions containing little or no cycloplegic agent areshown in Table 10 below.

TABLE 10 Compositions containing little or no cycloplegic agent #L1 #L2#L3 #L4 #L5 #L6 #L7 #L8 #L9 #L10 Aceclidine 1.75% 1.75% 1.75% 1.75%1.75% 1.75% 1.75% 1.75% 1.75% 1.75% Tropicamide 0.02% 0.02% 0.02% 0.02%0.02% — — — — — Mannitol 2.5% 2.5% 2.5% 2.5% 2.5% 2.5% 2.5% 2.5% 2.5%2.5% Polysorbate 80 0.75% 0.25% 0.25% 0.1% 0.1% 0.5%* 0.25% 0.25% 0.1%0.1% Carbopol® 940 0.95% 0.95% 0.95% 0.9% 0.95% 0.95%* 0.95%* 0.95%*0.9%* 0.95%* or CMC Glycerine Phosphate buffer 3 mM — 3 mM 3 mM 3 mM 3mM — 3 mM 3 mM 3 mM NaCl 0.5% 0.1% 0.05% — 0.1% 0.5%* 0.1% 0.05% — 0.1%Boric acid — 0.12% 0.2% 0.2% 0.12% — 0.12% 0.2% 0.2% 0.12% BAK 0.015%0.01% 0.01% 0.05% 0.01% 0.015% 0.01% 0.01% 0.05% 0.01% pH 5.0 5.0 5.05.0 5.0 5.0 5.0 5.0 5.0 5.0 #L11 #L12 #L13 #L14 #L15 #L16 #L17 #L18 #L19#L20 #L21 Aceclidine 1.65% 1.65% 1.75% 1.75% 1.65% 1.75% 1.75% 1.75%1.75% 1.75% 1.75% Tropicamide 0.01% — — 0.025% 0.025% 0.025% 0.025%0.025% 0.025% 0.015% 0.015% Mannitol 2.5% 2.5% 2.5% 2.5% 2.5% 2.5% 2.5%2.5% 2.5% 2.5% 2.5% Polysorbate 80 2% 2% 1% 0.10% 2.50% 2.50% 3.00%2.50% 2.50% 2.50% 2.50% Carbopol® 940 0.75% 0.75% 0.75% 0.75% 0.75%0.75% 0.75% 1.50% 0.75% 0.75% 0.75% or CMC Glycerine 0.10% 0.10% 0.10%0.10% 0.10% 0.10% 0.10% 0.10% 0.20% 0.20% 0.20% Phosphate buffer 3 mM 3mM 3 mM 3 mM 3 mM 3 mM 3 mM 3 mM 3 mM 3 mM 3 mM NaCl — — — — — — — — — —— Boric acid — — — — — — — — — — — BAK 0.01% 0.01% 0.01% 0.01% 0.015%0.015% 0.015% 0.015% 0.015% 0.015% 0.015% pH 5.0 5.0 5.0 5.0 5.25 5.255.25 5.25 5.25 5.25 5.25 #L22 #L23 #L24 #L25 #L26 #L27 #L28 #L29 #L30#L31 #L32 Aceclidine 1.65% 1.75% 1.75% 1.75% 1.75% 1.65% 1.75% 1.75%1.75% 1.75% 1.75% Tropicamide 0.025% 0.275% 0.020% 0.015% 0.027% 0.0275%0.0275% 0.0275% 0.025% 0.022% 0.0175% Mannitol 2.5% 2.5% 2.5% 2.5% 2.5%2.5% 2.5% 2.5% 2.5% 2.5% 2.5% Polysorbate 80 5% 5% 5% 5% 5% 5% 5% 5% 5%5% 5% Carbopol® 940 1.25% 1.45% 1.45% 1.45% 1.45% 1.25% 1.40% 1.40%1.50% 1.40% 1.50% or CMC Glycerine Phosphate buffer 3 mM 3 mM 3 mM 3 mM3 mM 3 mM 3 mM 3 mM 3 mM 3 mM 3 mM NaCl — — — — — — — — — — — Boric acid— — — — — — — — — — — BAK 0.01% 0.01% 0.01% 0.01% 0.01% 0.01% 0.01%0.01% 0.01% 0.01% 0.01% pH 5.0 5.0 5.0 5.0 6.0 5.0 5.0 5.0 5.0 5.0 6.0Pupil Size (mm) Reading vs. 3+ 3+ 3+ 3+ 3+ 3+ 3+ 3+ 3+ Baseline 40 cmDuration (hours) 7 10+ 10+ 10+ 10+ 7.0 10+ 10+ 10+ Ciliary Spasms 0.0 tr0.5 1.0 1.0 0.0 tr 0.5 1.0 1.0 Stinging 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.50.5 Blur (min) 1 1 1 1 1 1 1 1 1 Distance Blur Onset (min) 20 20 20 2020 20 20 20 20 Redness 1hr (0-4) 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5Redness 4hr (0-4) Overall Comfort sl sticky sl sticky sl sticky slsticky sl sticky sl sticky sl sticky sl sticky sl sticky OsmolarityEfficacy index: read*dur OVERALL (1-5) best best best best best bestbest best best #L33 #L34 #L35 #L36 #L37 #L38 #L47 #L48 #L49 #L50 #L51Aceclidine 1.75% 1.40% 1.40% 1.25% 1.45% 1.45% 1.45% 1.55% 1.65% 1.75%1.65% Tropicamide — — — — — 0.0200% — 0.0200% 0.0300% 0.0300% 0.0200%Brimonidine — — — — — — — — — — — Mannitol — — — — — — 2.5% 4.0% 2.5%2.5% 2.5% Polysorbate 80 — — — — — — — — — 5.00% — Polyoxy1 40 Stearate— — — — 5.5% 5.5% 5.5% 5.5% 5.5% — 5.5% Citrate 0.10% 0.10% 0.10% 0.10%0.10% 0.10% 0.10% 0.10% 0.10% — 0.10% Glycerine — 0.10% 0.10% 0.10%0.10% 0.10% 0.10% 0.10% 0.10% 0.10% 0.10% CMC — 1.45% 0.75% — 0.85%0.75% 0.75% 0.75% 0.75% 0.75% 0.75% HPMC — — — — — — — — — — — Carbopol®940 — — — — — — — — — — — NaCl 0.75% 0.75% 0.50% 0.50% 0.50% 0.50% 0.50%0.50% 0.50% 0.50% 0.00% Boric Acid — — — — — — — — — — — PostassiumBorate — — — — — — — — — — — Phosphate buffer 3 3 3 3 3 3 3 3 3 3 3Acetate — — — — — — — — — — — pH 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.255.0 5.0 BAK 0.015% 0.015% 0.015% 0.015% 0.015% 0.015% 0.015% 0.015%0.015% 0.015% 0.015% Pupil Size (mm) Reading vs. 3 3.25 3 2 3 2.5 1.50.5 1.5 1.5 1 Baseline 40 cm Duration (hours) 4 7 4.5 6.5 6 3 2 4 4 2Ciliary Spasms 4 4 3 2 3 2 0.5 0.5 0.5 0.5 0.5 Stinging 1.0 1.0 1.0 1.01 1 0.5 1 1 Blur (min) Distance blur none none none none none none nonenone none none none Onset (min) 20-11 20-12 20-13 20-14 20-15 20-1620-25 20-25 20-25 20-25 20-25 Redness Ihr (0-4) 2.0 1.5 0.5 0.5 0.5 0.50.5 0.5 0.5 Redness 4hr (0-4) Overall comfort poor poor poor fair poorpoor good good good good good Osmolarity hi hi hi hi hi hi hi hi hi hihi Efficacy index: 12 23 14 0 20 15 5 1 6 6 2 read*dur OVERALL (1-5) *** 1/2 * — * ** *** * * * * #L52 #L53 #L54 #L55 #L56 #L57 #L58 #L59 #L60#L61 #L62 Aceclidine 1.65% 1.65% 1.65% 1.75% 1.75% 1.65% 1.65% 1.65%1.65% 1.65% 1.75% Tropicamide 0.0100% 0.0250% 0.0000% 0.0000% 0.0250%0.0250% 0.0250% 0.0150% 0.0400% 0.0250% 0.0300% Brimonidine — — — — — —— — — — — Mannitol 2.5% 2.5% 2.5% 2.5% 2.5% 2.5% 2.5% 2.5% 2.5% 2.5%2.5% Polysorbate 80 2.00% 2.50% 2.00% 1.00% 0.10% 2.00% 2.50% 2.50%3.50% 2.50% 3.50% Polyoxy1 40 Stearate — — — — — — — — — — — Citrate —0.10% — — — — — — — — — Glycerine 0.10% 0.10% 0.10% 0.10% 0.10% 0.10%0.10% 0.10% 0.10% 0.10% 0.10% CMC 0.75% 0.85% 0.85% 0.85% 0.85% 0.85%0.85% 0.75% 0.60% 1.60% 0.60% HPMC — — — — — — — — — — — Carbopol® 940 —— — — — — — 0.75% 0.60% 0.60% NaCI 0.50% 0.50% — — — — — — — — — BoricAcid — — — — — — — — — — — Postassium Borate — — — — — — — — — — —Phosphate buffer 3 3 3 3 3 3 3 3 3 3 3 Acetate — — — — — — — — — — — pH5.0 5.0 5.0 5.3 5.3 5.3 5.00 5.00 5.00 5.00 5.00 BAK 0.015% 0.015%0.015% 0.015% 0.015% 0.015% 0.015% 0.015% 0.015% 0.015% 0.015% PupilSize (mm) Reading vs. 2.5 3 3 2 1.5 2.5 3 2 1.5 2.5 2.5 Baseline 40 cmDuration (hours) 6 5 6 4 4 6 5.5 7 3 7 7 Ciliary Spasms 0.5 0.5 2 2 0 00 0.5 0 0.5 0.5 Stinging 1 0.5 1 0.5 0.5 0.5 0.5 0.5 0,25 0.25 0,25 Blur(min) 1.5 1 2 Distance blur none none none none none none none none nonenone none Onset (min) 20-25 20-25 20-25 20-25 20-25 20-25 20-25 20-2520-25 20-25 20-25 Redness 1hr (0-4) 0.5 0.5 0.5 0.5 1.0 0.5 0.5 0.5 0.50.5 Redness 4hr (0-4) Overall comfort poor poor poor poor good good goodgood good good good Osmolarity hi hi nl nl n nl nl nl nl nl nl Efficacyindex: 15 15 18 8 6 15 17 14 5 18 18 read*dur OVERALL (1-5) ** **** ****#L63 #L64 #L65 #L66 #L67 #L68 #L69 #L70 #L71 #L72 #L73 Aceclidine 1.65%1.75% 1.65% 1.75% 1.75% 1.75% 1.75% 1.75% 1.75% 1.75% 1.75% Tropicamide0.0250% 0.0250% 0.0250% 0.0275% 0.0275% 0.0275% 0.0250% 0.0180% 0.0160%0.0160% 0.0150% Brimonidine — — — — — — — — — — — Mannitol 2.5% 2.5%2.5% 2.5% 2.5% 2.5% 2.5% 2.5% 2.5% 2.5% 2.5% Polysorbate 80 2.50% 3.50%4.00% 5.00% 5.00% 2.00% 2.00% 2.00% 2.25% 4.00% 4.00% Polyoxy1 40Stearate — — — — — — — — — — — Citrate — — — — — — — — — — — Glycerine0.10% 0.10% 0.10% — — — — — — — — CMC 0.75% 0.50% 0.75% — 1.35% 1.35%1.45% 1.45% 1.45% 1.45% HPMC — — — — — — — — — — — Carbopol® 940 0.50%1.35% — 1.45% — — — — — NaCl — — — — — — — — — — — Boric Acid — — — — —— — — — — — Postassium Borate — — — — — — — — — — — Phosphate buffer 3 33 3 3 3 3 3 3 3 3 Acetate — — — — — — — pH 5.00 5.00 5.00 5.0 5.0 5.05.0 5.0 5.0 5.0 5.0 BAK 0.015% 0.015% 0.015% 0.015% 0.015% 0.015% 0.015%0.015% 0.01% 0.01% 0.01% Pupil Size (mm) Reading vs. 2 2.5 2 2.75 2.752.75 2.75 2.75 2.75 2.75 3 Baseline 40 cm Duration (hours) 4 5 7 7 5.5 67 7 7 7.5 Ciliary Spasms 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5Stinging 0.5 0.5 0.5 0.5 0.5 0.5 Blur (min) 1 Distance blur none nonenone none none none none none none none none Onset (min) 20-25 20-2520-25 20-25 20-25 20-25 20-25 20-25 20-25 20-25 20-25 Redness 1hr (0-4)0.5 0.5 0.5 0.5 0.5 0.5 Redness 4hr (0-4) Overall comfort good good goodgood good good good good good good good Osmolarity nl nl nl nl nl nl nlnl nl nl nl Efficacy index: 8 0 10 19 19 15 17 19 19 19 23 read*durOVERALL (1-5) poor ? ** 1/2 ** **** **** _(1/2) #L74 #L75 #L76 #L77 #L78#L79 #L80 #L81 #L82 #L83 #L84 Aceclidine 1.75% 1.75% 1.75% 1.75% 1.75%1.75% 1.75% 1.75% 1.75% 1.65% 1.40% Tropicamide 0.0150% 0.0150% 0.0120%0.0110% 0.0100% 0.0000% — 0.0100% 0.0150% 0.0000% 0.0000% Brimonidine —— — — — 0.015% — — — — — Mannitol 2.5% 2.5% 2.5% 2.5% 2.5% 2.5% 2.5%2.5% 2.5% 2.5% 2.5% Polysorbate 80 5.00% 5.00% 5.00% 5.00% 5.00% 5.00% -6.00% 7.00% 0.00% 0.00% Polyoxy1 40 Stearate — — — — — — — — — — 5.5%Citrate — — — — — — — — — — — Glycerine — — — — — — — — — — — CMC 1.45%1.43% 1.43% 1.40% 1.40% 1.40% 1.40% 1.40% 1.40% 0.00% 0.75% HPMC — — — —— — — — — — — Carbopol® 940 — — — — — — — — — — — NaCl — — — — — — — —0.50% — — Boric Acid — — — — — — — — — — — Postassium Borate — — — — — —— — — — — Phosphate buffer 3 3 3 3 3 3 3 3 3 3 3 Acetate — — — — — — — —— — — pH 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 BAK 0.01% 0.01% 0.01%0.01% 0.01% 0.01% — 0.01% 0.01% 0.010% 0.010% Pupil Size (mm) Readingvs. 3.25 3.25 3.5 3.5 3.75 2.5 2.5 2.75 2.5 1 1.5 Baseline 40 cmDuration (hours) 7.5 7.5 7 8 9 8 7 5.5 5 3 3.5 Ciliary Spasms 0.5 0.5 11 1 2 2 0.5 0.5 1 1 Stinging 1.0 Blur (min) 1.5 1.5 1.5 Distance blurnone none none none none none none none none none none Onset (min) 20-2520-25 20-25 20-25 20-25 20-25 20-25 20-25 20-25 20-25 20-25 Redness 1hr(0-4) 2.0 Redness 4hr (0-4) Overall comfort good-exc good-exc good-excexc exc fair fair good good fair Osmolarity nl nl nl nl nl nl nl nl hinl nl Efficacy index: read*dur 24 24 25 28 34 20 18 15 13 3 5 OVERALL(1-5) ***** ***** *****! *****!! *****!! **** **** *** *** * #L85 #L86#L87 #L88 #L89 #L90 #L91 #L92 #L93 #L94 Aceclidine 1.75% 1.75% 1.75%1.75% 1.75% 1.75% 1.75% 1.75% 1.75% 1.75% Tropicamide 0.0000% 0.0100%0.0900% 0.0060% 0.0060% 0.0100% 0.0060% 0.0060% 0.0060% 0.0060%Brimonidine — — — — — — — — — — Mannitol 2.5% 2.5% 2.5% 2.5% 2.5% 2.5%2.5% 2.5% — 2.5% Polysorbate 80 5.00% 2.5% 2.5% 2.5% 2.5% 2.50% 2.50%2.75% 2.75% 3.50% Polyoxy1 40 Stearate — — — — — — — — — — Citrate — — —— — — — — — — Glycerine — — — — — — — — — — CMC 1.40% — — — — — — — — —HPMC — 1.75% 1.75% 1.75% 1.75% — — — — — Carbopol® 940 — — — — — 1.75%1.75% 1.80% 1.80% 1.80% NaCl 0.00% 0.50% — 0.50% — 0.50% 0.50% 0.50%0.50% 0.50% Boric Acid — — 0.35% — — — — — 0.25% — Postassium Borate — —0.47% — — — — — 0.37% — Phosphate buffer 3 3 3 3 4 3 3 3 3 3 Acetate — —— — — — — pH 5.0 5.0 5.0 5.0 6.0 5.0 5.0 5.0 5.0 5.0 BAK 0.010% 0.020%0.020% 0.020% 0.020% 0.02% 0.02% 0.02% 0.02% 0.02% Pupil Size (mm)Reading vs. 3.5 3.5 3.5 3.75 3.75 3.5 3.75 3.75 3.75 3.75 Baseline 40 cmDuration (hours) 7 8 7 9 9 7 7 8 8 8.5 Ciliary Spasms 2 0.5 0.5 0.5 0.50.5 0.5 0 0 0 Stinging 0.5 Blur (min) Distance blur 2.0 none none nonenone none none none none none Onset (min) 20-25 20-25 20-25 20-25 20-2520-25 20-25 20-25 20-25 20-25 Redness 1hr (0-4) 1.0 1.0 0.5 0.5 0.5 0.50.5 0.5 0.5 0.5 Redness 4hr (0-4) Overall comfort good good good goodgood Osmolarity nl nl nl nl 10 Efficacy index: 25 28 25 34 34 25 26 3030 32 read*dur OVERALL (1-5) **** ***** ***** ***** ***** ***** Allconcentration in weight by volume. mm denotes millimeters. cm denotescentimeters. min denotes minutes. %* denotes amount can optionally varyfrom about 0.01% to about 1% w/v. # denotes formulation can includepolysorbate 80 or not include polysorbate 80. Ciliary spasms scorescorrespond to the following: 0 = no discomfort; 0.5 = slight sting; 1 =noticeable squeeze/discomfort; 2 = pain for less than 30 minutes; 3 =pain for 1 hour or more; and 4 = severe to intolerable pain.

The efficacy index is demonstrated in FIG. 3 . In brief, the score iscalculated by multiplying the lines of improvement in near visual acuityby the number of hours the improvement lasts. For example, a score of: 5is equal to +1 lines of improvement in near visual acuity for 5 hours;10 is equal to +1.5 lines of improvement for 6.7 hours; 15 is equal to 2lines of improvement for 7.5 hours; 20 is equal to 2.5 lines ofimprovement for 8 hours; 25 is equal to 3+ lines of improvement for 8.3hours and 35 is equal to 3.75+ lines of improvement for 9 hours.

As demonstrated by comparing the Reading vs. Baseline at 40 cm andEfficacy Indexes of formulas #L33-#L37, formulas containing 1.40% ormore aceclidine are better at correcting presbyopia than those formulascontaining 1.25% aceclidine. Inversely, the lower concentration ofaceclidine results in better overall comfort to the user. The additionof 2.5% mannitol to formulas with 1.45% aceclidine improves overallcomfort but at the expense of reducing the presbyopic correcting effect(compare #L37 with #L47.) This reduction in near vision improvement isexacerbated with the addition of 4.0% mannitol (compare #L47 with #L48.)Increasing aceclidine concentrations to 1.65% or 1.75% overcome thereduction in near vision improvement seen with the addition of mannitol(compare #L47 with #L49 and #L50.)

Further, formulas containing 1.75% aceclidine and 2.5% mannitol have anincreased efficacy and duration in treating presbyopia that iscorrelated with an increase in polysorbate 80 up to 5.0% and theninversely correlated with a decrease in CMC from 1.45% to 1.40% (compareformulas #L66 to #L78.) Optimal formulations are demonstrated by #L77,#L78 and #L85-#L94, which each have the highest improve reading at 40 cmat between 3.5 and 3.75 visual acuity lines and the highest EfficacyIndex scores of 25 to 34, and the longest duration from 7 to 9 hours.The increase in effectiveness and duration of formulas from #L66 to #L78are also inversely correlated with a decrease in tropicamide from0.0275% to 0.01%. This same trend is demonstrated by the increase ineffectiveness (i.e. Reading vs. Baseline 40 cm) when comparing #L85through #L94.

This data demonstrates that mannitol can effectively reduce ciliaryspasms caused by aceclidine, thus reducing the need for a cycloplegicagent such as tropicamide. Further, this data demonstrates that theaddition of a non-ionic surfactant and viscosity agent can furtherenhance the efficacy and duration of compositions containing aceclidine,mannitol and low tropicamide.

This data also demonstrates that the use of a cycloplegic agent inaceclidine compositions containing polysorbate 80 and CMC is mostbeneficial to presbyopic correction when the cycloplegic agent is closerto 0.006% than 0.025%. Finally, this data demonstrates that compositionscomprising aceclidine and mannitol are sufficient to correct presbyopiawith tolerable pain.

Example 12 Use of Further High Tropicamide Formulations

The following examples are of aceclidine formulations containing morethan 0.0300 tropicamide.

TABLE 11 High tropicamide formulations #L39 #L40 #L41 #L42 #L43 #L44#L45 #L46 Aceclidine 1.45% 1.45% 1.40% 1.40% 1.40% 1.40% 1.40% 1.40%Tropicamide 0.035% 0.037% 0.040% 0.050% 0.055% 0.06% 0.08% 0.04%Polyoxy1 40 5.5% 5.5% 5.5% 5.5% 5.5% 5.5% 5.5% 5.5% Stearate Citrate0.10% 0.10% 0.10% 0.10% 0.10% 0.10% 0.10% 0.10% Glycerine 0.10% 0.10%0.10% 0.10% 0.10% 0.10% 0.10% 0.10% CMC 0.75% 0.75% 0.75% 0.75% 0.75%0.75% 0.75% 0.75% NaCl 0.50% 0.50% 0.50% 0.50% 0.50% 0.50% 0.50% 0.50%Phosphate buffer 3 3 3 3 3 3 3 3 pH 5.0 5.0 5.0 5.25 5.5 5.25 5.0 5.0BAK 0.015% 0.015% 0.015% 0.015% 0.015% 0.015% 0.015% 0.015% Reading vs.3.5 3.5 3.5 2 1 1 1 3 Baseline 40 cm Duration (hours) 6 6 6 2 2 1 1 6Ciliary Spasm 1 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Stinging 1.0 1.0 1.0 0.50.25 0.5 1 1 Distance blur none none none none none none none none Onset(min) 20-17 20-18 20-19 20-20 20-21 20-22 20-23 20-24 Redness 1hr (0-4)0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Overall comfort fair good good good goodgood good good Osmolarity hi hi hi hi hi hi hi hi Efficacy index: 21 2121 4 2 1 1 18 read*dur OVERALL (1-5) ** *** *** * 1/2* 1/4* 1/4* ***Ciliary spasms scores correspond to the following: 0 = no discomfort;0.5 = slight sting; 1 = noticeable squeeze/discomfort; 2 = pain for lessthan 30 minutes; 3-pain for 1 hour or more; and 4 = severe tointolerable pain.

As demonstrated by formulas #L39-#L41 and compared to formulas #L74-#L78in Table 10, formulas containing about 1.40% to about 1.45% aceclidine,about 0.035% to about 0.04% tropicamide, about 5.5% polyoxyl 40 stearateand about 0.75% CMC are almost, but not quite as effective at treatingpresbyopia as formulas containing about 1.65% to about 1.75% aceclidine,about 2.5% mannitol, about 5% polysorbate 80, about 1.40% CMC formulas.This effectiveness decreases dramatically when tropicamide is increasedto about 0.05% to about 0.08% tropicamide.

Example 13. Use of a Compound Containing Mannitol Formulation:

-   -   aceclidine 1.75% w/v    -   tropicamide 0.006% w/v    -   mannitol 2.5% w/v    -   polysorbate 80 2.75% w/v    -   NaCl 0.5% w/v    -   hydroxypropylmethyl cellulose 0.5%-1.80% w/v    -   phosphate buffer 3 mM    -   pH 5.0, and    -   BAK 0.020% as preservative.

Method:

The subject instilled 2 drops of the above formulation in each eye andthe excess wiped from lids and lashes.

Results:

Within 20 minutes, near vision improvement of about 3 lines of visualacuity was noted with very slight dimming. Throughout the day nearvision remained enhanced with no loss of distance vision. Further, ifthe subject previously suffered from any mild refractive errors distancevision was improved. Over a 5-8 hour period the pupil begins to slightlyrecover, and after a few hours the minimal dimming was no longer noted.Both excellent near vision near onset, and possibly still slightlyimproved near vision continued as the pupil slightly begins to increasefrom its minimal size earlier in the day.

Example 14. Use of a Preferred Embodiment Optimizing Tropicamide andHydroxypropyl Methyl Cellulose

Composition

Aceclidine 1.75% w/v Tropicamide 0.010% w/v  Mannitol 2.50% w/vPolysorbate 80 3.50% w/v NaCl 0.50% w/v HPMC 1.25% w/v BAK 0.02% w/vPhosphate buffer 3 mM pH 5.00

Method

The subject instilled 2 drops of the above formulation in each eye as 1single drop each eye and a second drop after 5 minutes.

Results:

Comfort, duration and efficacy were assessed. Stinging upon instillationand over the first hour was minimal with a score of 0.25 out of 4.Redness over the first hour was also minimal with a score of 0.5 out of4 assessed at 20 minutes. Onset of vision improvement occurred with thefirst 20 to 25 minutes after instillation. Baseline near vision (i.e. 40centimeters) was improved by 3.5 lines of visual acuity. Improvement innear vision lasted for 8.5 hours. Comparing this formula to those inTable 10, the Efficacy Index score was 29.75. Substituting HPMC 1.80%w/v with HPMC 1.65% w/v resulted in a slight reduction in near visionimprovement to 3.25 lines of visual acuity and a slight reduction induration to just over about 6 hour. Comparing this formula to those inTable 10, the Efficacy Index score was 19.5.

Example 15. Use of a Compound Containing Mannitol with Various NonionicSurfactants Compositions

Table 12 lists the active ingredients, excipients and theirconcentrations for compositions with both tested and prophetic examplesof nonionic surfactants.

Methods

The subject independently instilled 2 drops of the above compositions ineach eye and the excess wiped from lids and lashes.

Results

All nonionic surfactants tested demonstrate substantial near visionimprovement. Of those tested only Brij® 35 was marginal due to thesignificant corneal irritation, hyperemia and reduced duration thatresulted. Polysorbate 80 and poly 35 castor oil were most preferred,polyoxyl 40 stearate and poloxamer 407 excellent as well. However,polyoxyl 40 stearate caused a precipitate reaction with celluloseviscosity agents and added other stability issues.

Comfort and duration for each non-ionic surfactant were also tested andare noted in Table 12. Stinging and Redness are based on a scale of 0 to4 with 0 being none and 4 being the most severe. Other than Brij® 35stinging and redness were mild to nearly absent. Duration was excellentfor each nonionic surfactant tested.

TABLE 12 Comparing efficacy and comfort of various nonionic surfactantsPolyoxyl 35 Polyoxyl Polysorbate Poloxamer Solulan Polysorbate castor 40Poloxamer Brij® Tyloxapol 20 188 C-24 % w/v 80 oil stearate 407 35(prophetic) (prophetic) (prophetic) (prophetic) Aceclidine 1.75% 1.75%1.75% 1.75% 1.75% 1.75% 1.75% 1.75% 1.75% Tropicamide 0.006% 0.006%0.005% 0.005% 0.005% 0.006% 0.006% 0.006% 0.006% Mannitol 2.5% 2.5% 2.5%2.5% 2.5% 2.5% 2.5% 2.5% 2.5% Nonionic surfactant 3.5% 3.5% 3.5% 3.5%3.5% 3.5% 3.5% 3.5% 3.5% NaCl 0.50% 0.50% 0.50% 0.50% 0.50% 0.50% 0.50%0.50% 0.50% HPMC 1.80% 1.80% 1.80% 1.80% 1.80% 1.80% 1.80% 1.80% 1.80%BAK 0.02% 0.02% 0.02% 0.02% 0.02% 0.02% 0.02% 0.02% 0.02% Phosphatebuffer 3 mM 3 mM 3 mM 3 mM 3 mM 3 mM 3 mM 3 mM 3 mM pH 5.00 5.00 5.005.00 5.00 5.00 5.00 5.00 5.00 Stinging 0.25 0 0.5 0.5 2 0-2 0-2 0-2 0-2Redness 1hr 0.5 0.25 0.75 1 2.5 0.25- 0.25- 0.25- 0.25- 2.5 2.5 2.5 2.5Reading vs. 3.75 3.5 3 3 2 2-3.5 2-3.5 2-3.5 2-3.5 Baseline (40 cm)Duration (hours) 10 9 7 7 4 4-8 4-8 4-8 4-8 Efficacy Index 37.5 31.5 2121 8 8-37.5 8-37.5 8-37.5 8-37.5 read*dur Onset (min) 20-25 20-25 20-2520-25 30-40 20-40 20-40 20-40 20-40

Example 16. Use of a Compound Containing Optimizing Nonionic Surfactantand Antioxidant Additives and Concentrations Compositions

Aceclidine 1.75% w/v Tropicamide 0.010% w/v  Mannitol 2.50% w/vPolysorbate 80 4.00% w/v NaCl 0.00% w/v HPMC 1.25% w/v (high MW equalingviscosity of about 400 cps units) BAK 0.02% w/v Sorbate 0.12% w/v BAK0.02% w/v EDTA 0.01% Citrate buffer 3 mM pH 5.00

Method

2 subjects instilled 2 drops each of the above formulation in each eyeabout 5 minutes apart.

Results:

Comfort, duration and efficacy were assessed. Stinging upon instillationand over the first hour was minimal for each subject with a score of0.50 out of 4 for about 15 seconds. Redness over the first hour was alsominimal for each subject with a score of 0.25 out of 4 assessed at 20minutes. Onset of vision improvement occurred with the first 20 to 25minutes after instillation. For subject 1 baseline near vision (i.e. 40centimeters) was improved by 4.0-4.25 lines of visual acuity and lastedfor 11.5 hours. For subject 2 baseline near vision was improved by 3.5lines of visual acuity and lasted for 9.5 hours. The Efficacy Indexscore was 47.38 and 33.25, among the highest achieved for anyformulation.

Example 17. Aceclidine to Treat Presbyopia Method

A 1.75% aceclidine in saline solution was formulated. One drop wasinstilled into each eye of a presbyopic subject. The visual acuity ofthe subject was tested both before and after instillation using a LogMARchart.

Results

The subject recorded a best corrected distance visual acuity of LogMAR0.50 at 45 centimeters near test card and LogMAR −0.12 best correcteddistance acuity at 1-meter distance prior to instillation. Afterinstillation the subject recorded a LogMAR score of 0.22 at 45centimeters distance and maintained a −0.12 LogMAR score at 1-meterdistance. This improved reading distance acuity was maintained for 3.5hours post instillation. Noticeable degradation in reading distanceacuity began at 4 hours post instillation.

What is claimed is:
 1. A cold chain storage system comprising (i) anopthalmological composition comprising aceclidine in a stable solution,wherein the aceclidine remains in solution when the solution ismaintained at about 2° C. to about 8° C.; and (ii) a container suitablefor cold chain storage.
 2. The system of claim 1, wherein degradation ofthe aceclidine is inhibited in the stable solution.
 3. The system ofclaim 1, wherein the opthalmological composition further comprises aviscosity enhancer.
 4. The system of claim 3, wherein the viscosity ofthe composition is from about 1 to about 5,000 centipoise in storage. 5.The system of claim 1, wherein the opthalmological composition furthercomprises a surfactant selected from the group consisting of an anionicsurfactant, a nonionic surfactant, and a combination thereof.
 6. Thesystem of claim 5, wherein the surfactant is a nonionic surfactant. 7.The system of claim 1, wherein the opthalmological composition comprisesa pH of about 4.0 to about 8.0.
 8. The system of claim 7, wherein theopthalmological composition comprises a pH of about 4.0 to about 6.0. 9.The system of claim 1, wherein the viscosity of the opthalmologicalcomposition is above 20 cps at 1/s shear upon topical application. 10.The system of claim 1, wherein at least 88% of the aceclidine remains insolution after 1 to 3 months when the solution is maintained at about 2°C. to about 8° C.
 11. A method of treating presbyopia in a subject inneed thereof, the method comprising: a. providing an opthalmologicalcomposition comprising aceclidine in a stable solution, wherein theaceclidine remains in solution when the solution is maintained at about2° C. to about 8° C.; b. storing the opthalmological composition at atemperature from about 2° C. to about 8° C.; and c. administering theopthalmological composition to an eye of the subject.
 12. The method ofclaim 11, wherein the opthalmological composition is administered toboth eyes of the subject.
 13. The method of claim 12, whereinadministration to both eyes of the subject enhances near vision of thesubject greater than compared to administration to either eye of thesubject separately.
 14. The method of claim 11, wherein a size of apupil of the eye is reduced to from about 1.5 to about 2.5 millimeterssubsequent to administering.
 15. The method of claim 11, whereindegradation of the aceclidine is inhibited in the stable solution. 16.The method of claim 11, wherein the ophthalmological composition furthercomprises a viscosity enhancer.
 17. The method of claim 16, wherein theviscosity of the composition is from about 1 to about 5,000 centipoiseduring storing.
 18. The method of claim 11, wherein the opthalmologicalcomposition further comprises a surfactant selected from the groupconsisting of an anionic surfactant, a nonionic surfactant, and acombination thereof.
 19. The method of claim 18, wherein the surfactantis a nonionic surfactant.
 20. The method of claim 11, wherein theophthalmological composition comprises a pH of about 4.0 to about 8.0.21. The method of claim 20, wherein the opthalmological compositioncomprises a pH of about 4.0 to about 6.0.
 22. The method of claim 11,wherein the viscosity of the ophthalmological composition is above 20cps at 1/s shear during administering.
 23. The method of claim 11,wherein at least 88% of the aceclidine remains in solution after 1 to 3months when the solution is maintained at about 2° C. to about 8° C.